Note: Descriptions are shown in the official language in which they were submitted.
COMPOUNDS FOR TREATMENT OF CANCER
FIELD OF THE INVENTION
[001] The present invention relates to novel compounds having anti-cancer
activity, methods of
making these compounds, and their use for treating cancer, treating drug-
resistant tumors, drug-
resistant cancer, metastatic cancer, metastatic melanoma, drug resistant
melanoma, prostate cancer
and drug resistant prostate cancer.
BACKGROUND OF THE INVENTION
[002] Cancer is the second most common cause of death in the United States,
exceeded only by
heart disease. In the United States, cancer accounts for 1 of every 4 deaths.
The 5-year relative
.. survival rate for all cancer patients diagnosed in 1996-2003 is 66%, up
from 50% in 1975-1977
(Cancer Facts & Figures American Cancer Society: Atlanta, GA (2008)). This
improvement in
survival reflects progress in diagnosing at an earlier stage and improvements
in treatment.
Discovering highly effective anticancer agents with low toxicity is a primary
goal of cancer
research.
[003] Microtubules are cytoskeletal filaments consisting of 443-tubu1in
heterodimers and are .
involved in a wide range of cellular functions, including shape maintenance,
vesicle transport,
cell motility, and division. Tubulin is the major structural component of the
microtubules and a
well verified target for a variety of highly successful anti-cancer drugs.
Compounds that are able
to interfere with microtubule-tubulin equilibrium in cells are effective in
the treatment of cancers.
Anticancer drugs like taxol and vinblastine that are able to interfere with
microtubule-tubulin
equilibrium in cells are extensively used in cancer chemotherapy. There are
three major classes of
antimitotic agents. Microtubule-stabilizing agents, which bind to fully formed
microtubules and
prevent the depolymerization of tubulin subunits, are represented by taxanes
and epothilones.
The other two classes of agents are microtubule-destabilizing agents, which
bind to tubulin
dimers and inhibit their polymerization into microtubules. Vina alkaloids such
as vinblastine
bind to the vinca site and represent one of these classes. Colchicine and
colchicine-site binders
interact at a distinct site on tubulin and define the third class of
antimitotic agents.
[004] Both the taxanes and vinca alkaloids are widely used to treat human
cancers, while no
colchicine-site binders are currently approved for cancer chemotherapy yet.
However, colchicine
1
CA 3030689 2019-01-18
binding agents like combretastatin A-4 (CA-4) and ABT-751 (Figure 19), are now
under clinical
investigation as potential new chemotherapeutic agents (Luo, Y.; Hradil, V.
P.; Frost, D. J.;
Rosenberg, S. H.; Gordon, G. B.; Morgan, S. J.; Gagne, G. D.; Cox, B. F.;
Tahir, S. K.; Fox, G.
B., ABT-751, "A novel tubulin-binding agent, decreases tumor perfusion and
disrupts tumor
vasculature". Anticancer Drugs 2009, 20(6), 483-92.; Mauer, A. M.; Cohen, E.
E.; Ma, P. C.;
Kozloff, M. F.; Schwartzberg, L.; Coates, A. I.; Qian, J.; Hagey, A. E.;
Gordon, G. B., "A phase
II study of ABT-751 in patients with advanced non-small cell lung cancer". J
Thorac Oncol
2008, 3(6), 631-6.; Rustin, G. J.; Shreeves, G.; Nathan, P. D.; Gaya, A.;
Ganesan, T. S.; Wang,
D.; Boxall, J.; Poupard, L.; Chaplin, D. J.; Stratford, M. R.; Ballcissoon,
J.; Zweifel, M., "A Phase
lb trial of CA4P (combretastatin A-4 phosphate), cattoplatin, and paclitaxel
in patients with
advanced cancer". Br J Cancer 2010, 102(9), 1355-60.).
[005] Unfortunately, microtubule-interacting anticancer drugs in clinical use
share two major
problems, resistance and neurotoxicity. A common mechanism of multidrug
resistance (MDR),
namely ATP binding cassette (ABC) transporter protein-mediated drug efflux,
limits their
efficacy (Green, H.; Rosenberg, P.; Soderkvist, P.; Horvath, G.; Peterson, C.,
"beta-Tubulin
mutations in ovarian cancer using single strand conformation analysis-risk of
false positive
results from paraffin embedded tissues". Cancer Letters 2006, 236(1), 148-54.;
Wang, Y.;
Cabral, F., "Paclitaxel resistance in cells with reduced beta ¨tubulin"
Biochimica et Biophysica
Acta, Molecular Cell Research .2005, 1744(2), 245-255.; Leslie, E. M.; Deeley,
R. G.; Cole, S. P.
C., "Multidrug resistance proteins: role of P-glycoprotein, MRP1, MRP2, and
BCRP (ABCG2) in
tissue defense". Toxicology and Applied Pharmacology 2005,204(3), 216-237.).
[006] P-glycoproteins (P-gp, encoded' by the MDR1 gene) are important members
of the ABC
superfamily. P-gp prevents the intracellular accumulation of many cancer drugs
by increasing
their efflux out of cancer cells, as well as contributing to hepatic, renal,
or intestinal clearance
pathways. Attempts to co-administer P-gp modulators or inhibitors to increase
cellular
availability by blocking the actions of P-gp have met with limited success
(Gottesman, M. M.;
Pastan, L, "The multidrug transporter, a double-edged sword". J Biol Chem
1988, 263(25),
12163-6.; Fisher, G. A.; Sikic, B. I., "Clinical studies with modulators of
multidrug resistance".
Hematology/Oncology Clinics of North America 1995, 9(2), 363-82).
[007] The other major problem with taxanes, as with many biologically active
natural products,
is its lipophilicity and lack of solubility in aqueous systems. This leads to
the use of emulsifiers
2
CA 3030689 2019-01-18
like Cremophor EL and TweenTm 80 in clinical preparations. A number of
biologic effects related to
these drug formulation vehicles have been described, including acute
hypersensitivity reactions and
peripheral neuropathies (Hennenfent, K. L.; Govindan, R., "Novel formulations
of taxanes: a review.
Old wine in a new bottle?" Ann Oncol 2006, 17(5), 735-49.; ten Tije, A. J.;
Verweij, J.; Loos, W. J.;
Sparreboom, A., "Pharmacological effects of formulation vehicles: implications
for cancer
chemotherapy". Gin Pharmacokinet 2003, 42(7), 665-85.).
[008] Compared to compounds binding the paclitaxel- or vinca alkaloid binding
site, colchicine-
binding agents usually exhibit relatively simple structures. Thus providing a
better opportunity for
oral bioavailability via structural optimization to improve solubility and
pharmacokinetic (PK)
.. parameters. In addition, many of these drugs appear to circumvent P-gp-
mediated MDR. Therefore,
these novel colchicine binding site targeted compounds hold great promise as
therapeutic agents,
particularly since they have improved aqueous solubility and overcome P-gp
mediated MDR.
[009] Prostate cancer is one of the most frequently diagnosed noncutaneous
cancers among men in
the US and is the second most common cause of cancer deaths with over 180,000
new cases and
almost 29,000 deaths expected this year. Patients with advanced prostate
cancer undergo androgen
deprivation therapy (ADT), typically either by luteinizing hormone releasing
hormone (LHRH)
agonists or by bilateral orchiectomy. Androgen deprivation therapy not only
reduces testosterone, but
estrogen levels are also lower since estrogen is derived from the
aromatization of testosterone, which
levels are depleted by ADT. Androgen deprivation therapy-induced estrogen
deficiency causes
significant side effects which include hot flushes, gynecomastia and
mastalgia, bone loss, decreases
in bone quality and strength, osteoporosis and life-threatening fractures,
adverse lipid changes and
higher cardiovascular disease and myocardial infarction, and depression and
other mood changes.
[0010] Leuprolide acetate (Lupron8) is a synthetic nonapeptide analog of
naturally occurring
gonadotropin-releasing hormone (GnRH or LHRH). Leuprolide acetate is an LHRH
superagonist
that eventually suppresses LH secretion by the pituitary. Leuprolide acetate
acts as a potent inhibitor
of gonadotropin secretion, resulting in suppression of ovarian and testicular
steroidogenesis. In
humans, administration of leuprolide acetate results in an initial increase in
circulating levels of
luteinizing hormone (LH) and follicle stimulating hormone (FSH), leading to a
transient increase in
levels of the gonadal steroids (testosterone and dihydrotestosterone in
3
Date Recue/Date Received 2020-07-24
males, and estrone and estradiol in premenopausal. females). However,
continuous administration
of leuprolide acetate results in decreased levels of LH and FSH. In males,
testosterone is reduced
to castrate levels (below 50 ng,/dL). In premenopausal females, estrogens are
reduced to
postmenopausal levels. Testosterone is a known stimulus for cancerous cells of
the prostate.
Suppressing testosterone secretion or inhibiting the actions of testosterone
is thus a necessary
component of prostate cancer therapy. Leuprolide acetate can be used for LH
suppression, which
is the reduction and lowering of serum testosterone to castrate levels to
treat prostate cancer.
[0011] Malignant melanoma is the most dangerous form of skin cancer,
accounting for about
75% of skin cancer deaths. The incidence of melanoma is rising steadily in
Western populations.
The number of cases has doubled in the past 20 years. Around 160,000 new cases
of melanoma
are diagnosed worldwide each year, and it is more frequent in males and
Caucasians. According
to a WHO Report, about 48,000 melanoma-related deaths occur worldwide per
year.
[0012] Currently there is no effective way to treat metastatic melanoma. It is
highly resistant to
current chemotherapy, radiotherapy, and immunotherapy. Metastatic melanoma has
a very poor
prognosis, with a median survival rate of 6 months and a 5-year survival rate
of less than 5%. In
the past 30 years, dacarbazine (DTIC) is the only FDA-approved drug for
metastatic melanoma.
However, it provides only less than 5% of complete remission in patients. In
recent years, great
efforts have been attempted in fighting metastatic melanoma. Neither
combinations of DTIC with
other chemotherapy drugs (e.g., cisplatin, vinblastine, and carmustine) nor
adding interferon- a2b
to DTIC have shown a survival advantage over DTIC treatment alone. Most
recently, clinical
trials with antibodies and vaccines to treat metastatic melanoma also failed
to demonstrate
satisfactory efficacy.
[0013] Melanoma cells have low levels of spontaneous apoptosis in vivo
compared with other
tumor cell types, and they are relatively resistant to drug-induced apoptosis
in vitro. The natural
role of melanocytes is to protect inner organs from UV light, a potent DNA
damaging agent.
Therefore, it is not surprising that melanoma cells may have special DNA
damage repair systems
and enhanced survival properties. Moreover, recent studies showed that, during
melanoma
progression, it acquired complex genetic alterations that led to
hyperactivation of efflux pumps,
detoxification enzymes, and a multifactorial alteration of survival and
apoptotic pathways. All
these have been proposed to mediate the multidrug-resistant (MDR) phenotype of
melanoma.
With the rapidly rising incidence of this disease and the high resistance to
current therapeutic
4
CA 3030689 2019-01-18
agents, developing more effective drugs for advanced melanoma and other cancer
types that can
effectively circumvent MDR will provide significant benefits to cancer
patients.
SUMMARY OF THE INVENTION
[0014] In one aspect of the invention, there is provided a compound
represented by the structure of
formula XI:
X-4N 0
Me0 OMe
OMe (XI)
wherein
X is NH or S;
Q is 0, NH or S; and
A is substituted or unsubstituted single-, fused- or multiple-ring, aryl or
(hetero)cyclic
ring systems; substituted or unsubstituted, saturated or unsaturated N-
heterocycles;
substituted or unsubstituted, saturated or unsaturated S-heterocycles;
substituted or
unsubstituted, saturated or unsaturated 0-heterocycles; substituted or
unsubstituted,
saturated or unsaturated cyclic hydrocarbons; or substituted or unsubstituted
or saturated
or unsaturated mixed heterocycles;
wherein said A ring is optionally substituted by 1-5 substituents which are
independently
0-alkyl, 0-haloalkyl, F, Cl, Br, Iõhaloalkyl, CF3, CN, -CH2CN, NH2, hydroxyl, -
(CH2),NHCH3, -(CH2),NH2, -(CH2)1N(CH3)2, -0C(0)CF3, Ci-05 linear or branched
alkyl,
haloalkyl, alkylamino, aminoalkyl, -OCH,Ph, -NHCO-alkyl, COOH, -C(0)Ph, C(0)0-
alkyl, C(0)H, -C(0)NH2 or NO2; and
i is an integer between 0-5.
[0014a] In another aspect of the invention, there is provided a compound
represented by the structure of
formula XI:
CA 3030689 2019-01-18
0 ________________________________________ Xi To
Me0 OMe
OMe (XI)
wherein
X is a bond;
Q 0 or NH; and
A is a substituted phenyl, or a substituted or unsubstituted indole;
wherein said A ring is optionally substituted by 1-5 substituents which are
independently
0-alkyl, 0-haloalkyl, F, Cl, Br, I, haloalkyl, CF3, CN, -CII2CN, N112,
hydroxyl,
-(CH2)1NHCH3, -(CH2)1NH2, -(CH2)1N(CH3)2, -0C(0)CF3, CI-05 linear or branched
alkyl, haloalkyl, alkylamino, aminoalkyl, -OCH2Ph, -NHCO-alkyl, COOH, -C(0)Ph,
C(0)0-alkyl, C(0)H, -C(0)NH2 or NO2; and
i is an integer between 0-5.
[01413] In another aspect of the invention, there is provided a compound
represented by the
structure of formula XI:
0 ______________________________ x---4
Me0 OMe
OMe (XI)
wherein
X is a bond;
Q is NH; and
A is a substituted phenyl, or a substituted indole;
wherein said A ring is optionally substituted by 1-5 substituents which are
independently
0-alkyl, 0-haloalkyl, F, Cl, Br, I, haloalkyl, CF3, CN, -CH2CN, NH2, hydroxyl,
5a
CA 3030689 2019-01-18
-(CH2)iNHCH3, -(CH2)iNH2, -(CH2)iN(CH3)2, -0C(0)CF3, Ci-05 linear or branched
alkyl,
haloalkyl, alkylamino, aminoalkyl, -OCH2Ph, -NHCO-alkyl, COOH, -C(0)Ph, C(0)0-
alkyl,
C(0)H, -C(0)NH2 or NO2; and
i is an integer between 0-5;
or its isomer, pharmaceutically acceptable salt, pharmaceutical product,
tautomer, hydrate, N-
oxide, or combinations thereof.
[0014c] In another aspect, there is provided a compound represented by the
structure of formula XI:
A X--4
Me0
ome
(XI)
wherein
X is NH or a bond;
Q is NH or S; and
A is a substituted phenyl, or a substituted indole;
wherein said A ring is optionally substituted by 1-5 substituents which are
independently 0-
alkyl, 0-haloalkyl, F, Cl, Br, I, haloalkyl, CF3, CN, -CH2CN, NH2, hydroxyl,
-(CH2)iNHCH3, -(CH2)iNH2, -(CH2)iN(CH3)2, -0C(0)CF3, Ci-05 linear or branched
alkyl,
haloalkyl, alkylamino, aminoalkyl, -OCH2Ph, -NHCO-alkyl, COOH, -C(0)Ph, C(0)0-
alkyl,
C(0)H, -C(0)NH2 or NO2; and
i is an integer between 0-5;
provided if X is a bond, then the indole ring is not unsubstituted;
or its isomer, pharmaceutically acceptable salt, tautomer, hydrate, N-oxide,
or combinations
thereof.
[0015] In one embodiment, this invention is directed to a compound represented
by the structure of
formula VIII:
5b
Date Recue/Date Received 2020-07-24
OMe
0
OMe
N OMe
HN
(R4)11 R6
R5
(VIII)
R4, R5 and R6 are independently hydrogen, 0-alkyl, 0-haloalkyl, F, Cl, Br, I,
haloalkyl,
CF3, CN, -CH2CN, NH2, hydroxyl, -(CH2)1NHCH3, 4CH2)1NH2, -(CH2)1N(CH3)2, -
OC(0)CF3, C1-05 linear or branched alkyl, haloalkyl, alkylamino, aminoalkyl, -
OCH2Ph,
-NHCO-alkyl, COOH, -C(0)Ph, C(0)0-alkyl, C(0)H, -C(0)NH2 or NO2;
Q is S, 0 or NH;
i is an integer between 0-5; and
n is an integer between 1-3.
10010 In one embodiment, this invention is directed to a compound
represented by the structure
of formula XI(b):
= OMe
0
r(J
OMe
HN. N OMe
HN R5
(R4)n
NH
XI(b)
wherein R4 and R5 are independently hydrogen, 0-alkyl, 0-haloalkyl, F, Cl, Br,
I,
haloalkyl, CF3, CN, -CH2CN, NH2, hydroxyl, -(CH2)1NHCH3, -(CH2)1NI42, -
(CH2)1N(CH3)2, -0C(0)CF3, C1-05 linear or branched alkyl, haloalkyl,
alkylamino,
6
CA 3030689 2019-01-18
aminoalkyl, -OCH2Ph, -NHCO-alkyl, COOH, -C(0)Ph, C(0)0-alkyl, C(0)H, -C(0)NH2
or NO2;
i is an integer from 0-5; and
n is an integer between 1-4.
[0017] In one
embodiment, this invention is directed to a compound represented by the
structure
of formula XI(e):
OMe
0
OMe
S N OMe
-yr
HN R5
(R4)n
NH
XI(c)
wherein R4 and R5 are independently hydrogen, 0-alkyl, 0-haloalkyl, F, CI, Br,
I,
haloalkyl, CF3, CN, -CH2CN, NH2, hydroxyl, -(CH2),NHCH3, -(CH2)1NH2, -
(CH2),N(CH3)2, -0C(0)CF3, C1-05 linear or branched alkyl, haloalkyl,
alkylamino,
aminoalkyl, -OCH2Ph, -NHCO-alkyl, COOH, -C(0)Ph, C(0)0-alkyl, C(0)H, -C(0)NH2
or NO2;
i is an integer from 0-5; and
n is an integer between 1-4.
[0018] En one
embodiment, this invention is directed to a compound represented by the
structure of formula XI(e):
7
CA 3030689 2019-01-18
OMe
0
OMe
HN N OMe
R5
HN
(R4)n XI(e)
wherein R4 and R5 are independently hydrogen, 0-alkyl, 0-haloalkyl, F, Cl, Br,
I,
haloalkyl, CF3, CN, -CH2CN, NH2, hydroxyl, -(CH2)1NI-ICH3, -(CH2)1NH2, -
(CH2)1N(CH3)2, -0C(0)CF3, CI-Cs linear or branched alkyl, haloalkyl,
alkylamino,
aminoalkyl, -OCH2ph, -NHCO-alkyl, COOH, -C(0)Ph, C(0)0-alkyl, C(0)H, -C(0)NH2
or NO2;
i is an integer from 0-5; and
n is an integer between 1-4.
[0019] In another embodiment, this invention is directed to the following
compounds: (2-
(phenylamino)thiazol-4-y1)(3,4,5-trimethox yphenyl)methanone (5a), (2-(p- tol
ylami no)th azol-4-
yl)(3,4,5-trimethoxyphenyl)methanone (5b), (2-(p-
fluorophenylamino)thiaw1-4-y1)(3,4,5-
trimethox yphenyl)methanone (Sc), (2-(4-
chlorophenylarnino)thiazol-4-y1)(3,4,5-
trimethoxyphenyl)methanone (5d), (2-
(phenylamino)-1H-imidazol-4-y1)(3,4,5-
trimethoxyphenypmethanone (5e), 241H-
indo1-3-y1)-1H-imidazol-4-y1)(3,4,5-
trimethoxyphenyl)methanone (17ya); and (2-(1H-
indo1-5-ylamino)thiazol-4-y1)(3,4,5-
_
trimethoxyphenyl)methanone (55).
100201 In another embodiment, the compound of this invention is its isomer,
pharmaceutically
acceptable salt, pharmaceutical product, tautomer, hydrate, N-oxide, or
combinations thereof.
[0021] In one
embodiment, this invention is directed to a pharmaceutical composition
comprising a compound of this invention and a pharmaceutically acceptable
carrier.
[0022] In one
embodiment this invention is directed to a method of (a) treating,
suppressing,
reducing the severity, reducing the risk, or inhibiting cancer; (b) treating a
drug resistant tumor or
tumors; and (c) destroying a cancerous cell comprising administering a
compound of this invention.
8
CA 3030689 2019-01-18
In another embodiment the cancer is selected from the group consisting of
prostate cancer, breast
cancer, ovarian cancer, skin cancer, melanoma, lung cancer, colon cancer,
leukemia, renal
cancer, CNS cancer, and combinations thereof.
[0022a] In another aspect of the invention, there is provided use of a
compound as defined by
the invention for treatment of cancer, wherein said cancer is selected from
the group consisting
of prostate cancer, breast cancer, ovarian cancer, skin cancer, melanoma, lung
cancer, colon
cancer, leukemia, renal cancer, CNS cancer, and combinations thereof.
[0022b] In another aspect of the invention, there is provided use of a
compound as defined by
the invention for treatment of a drug resistant tumor, wherein said tumor is
selected from the
group consisting of melanoma cancer tumor, metastatic melanoma tumor, prostate
cancer tumor
and ovarian cancer tumor, and combinations thereof.
.. [0022c] In another aspect of the invention, there is provided use of a
compound as defined by
the invention in the preparation of a medicament for treatment of cancer,
wherein said cancer is
selected from the group consisting of prostate cancer, breast cancer, ovarian
cancer, skin cancer,
melanoma, lung cancer, colon cancer, leukemia, renal cancer, CNS cancer, and
combinations
thereof.
[0022d] In another aspect of the invention, there is provided use of a
compound as defined by
the invention in the preparation of a medicament for treatment of a drug
resistant tumor, wherein
said tumor is selected from the group consisting of melanoma cancer tumor,
metastatic
melanoma tumor, prostate cancer tumor and ovarian cancer tumor, and
combinations thereof.
[0022e] In another aspect of the invention, there is provided a compound as
defined by the
invention for use in treatment of cancer, wherein said cancer is selected from
the group
consisting of prostate cancer, breast cancer, ovarian cancer, skin cancer,
melanoma, lung cancer,
colon cancer, leukemia, renal cancer, CNS cancer, and combinations thereof.
9
CA 3030689 2019-01-18
[0022f] In another aspect of the invention, there is provided a compound as
defined by the
invention for use in treatment of a drug resistant tumor, wherein said tumor
is selected from the
group consisting of melanoma cancer tumor, metastatic melanoma tumor, prostate
cancer tumor
and ovarian cancer tumor, and combinations thereof
[0022g] In other aspects of the invention, there is provided use of a compound
of the invention
for treatment of prostate cancer or in the preparation of a medicament for
treatment of prostate
cancer, or a compound of the invention for use in treatment of prostate
cancer.
BRIEF DESCRIPTION OF THE DRAWINGS
[0023]
The subject matter regarded as the invention is particularly pointed out
and distinctly
claimed in the concluding portion of the specification. The invention,
however, both as to
organization and method of operation, together with objects, features, and
advantages thereof, may
best be understood by reference to the following detailed description when
read with the
accompanying drawings in which:
[0024]
Figure 1 depicts the synthesis of the diverse B-ring template: oxazole.
Reagents and
conditions: (a) Me0H, CH3C0C1, 83%; (b) Benzimidic acid ethyl ester, CH2C12,
Et3N, 96%; (c)
Li0H, Me0H, H20, 65%; (d) EDCI, HOBt, NMM, CH3OCH3NWHC1, 61%; (e) 3,4,5-
trimethoxyphenylmagnesium bromide, THF, 48%-71%; CBrC13, DBU, CH2C12, 56%.
[0025]
Figure 2 depicts the synthesis of the diverse B-ring templates. Reagents
and conditions:
(a) EDCI, HOBt, NMM, CH3OCH3NWHC1, CH2C12, 51-95%; (b) 3,4,5-trimethoxyphenyl-
magnesium bromide, THE, 48-78%; (c) LAH, -78 C, THE, 85%; (d) Dess-Martin
reagent, CH2C12,
81%; (e) EDCI, HOBt, NMM, 3,4,5-trimethoxybenzoic acid, CH2C12, 58%.
[0026]
Figure 3 depicts the synthetic scheme of compounds of this invention.
Reagents and
conditions: (a) Me0H/pH=6.4 phosphate buffer, RT; (b) EDCI, HOBt, NMM,
HNCH3OCH3; (c)
CBrC13, DBU, CH2C12; (d) 3,4,5-trimethoxyphenylmagnesium bromide, THF; (e)
isopropyl
triphenylphosphonium iodide, n-BuLi, THE;
LAH, THE; (g) For 2e-cis and 2e-trans,
NH2OH=HC1, C2H5OH, H20, NaOH; For 2g and 2h, NH20Me=HC1, pyridine; (h) TsCl,
NaH, basic
A1203; (i) NH2NH2.xH20, CH2C12, t-BuOH; (j) diethyl cyanomethylphosphonate, n-
BuLi, THE; (k)
bis-trimethylsilylcarbodiimide, TiC14, CH2C12; (1) EDCI, HOBt, Et3N, 3,4,5-
trimethoxyaniline,
CH2C12.
9a
Date Recue/Date Received 2020-07-27
[0027] Figure 4 depicts the synthetic scheme of compounds of this invention.
Reagents and
conditions: (a) bromine, Et0H; (b) benzothioamide, Et0H, reflux; (c) EDCI,
HOBt, NMM,
HNCH30CH3, CH2C12; (d) CBrC13, DBU, C112C12; (e) LAH, THF; (f) 5-(bromomethyl)-
1,2,3-
-
9b
CA 3030689 2019-01-18
trimethoxybenzene, Ph3P, THF; (g) n-BuLi, THF; (h) (1) HC1, H20; (2) NaNO2,
H20, 0 C; (i) ethyl
potassium xanthate; (j) KOH/Et0H; (k) H20, HC1; (1) 5-iodo-1,2,3-
trimethoxybenzene, Cul, 1-
13u0Na; (m) 2 equiv or 1 equiv m-CPBA, CH2C12; (n) 3,4,5-trimethoxyaniline,
NEt3, DMF.
[0028] Figure 5 depicts the synthetic scheme of compounds of this
invention. Reagents and
conditions: (a) L-cysteine, Et0H, 65 C; (b) EDCI, HOBt, NMM, HNCH3OCH3,
CH2C12; (c)
TBDMSC1, imidazole, THF; (d) 3,4,5-trimethoxyphenylbromide, BuLi, THF; (e)
TBAF, THF; (f)
SOCl2, Et20; (g) NH3, Me0H; (h) P0CI3; (i) PhS02C1, Bu4NHSO4, toluene, 50%
NaOH; (j) 1 N
NaOH, Et0H, reflux; (k) Boc20, 1 N NaOH, 1,4-dioxane; (1) CBrC13, DBU, CH2C12;
(m) 4 N Ha
in 1,4-dioxane; (n) Nan, DMF, Mel; (o) HCHO, NaBH3CN, Et3N.
[0029] Figure 6 depicts the synthetic scheme of compounds of this
invention. Reagents and
conditions: (a) Et0H, 65 C; (b) NaOH, C2H5OH, refluxing; (c) EDCI, HOBt, NMM,
HNCH3OCH3, CH2C12; (d) 3,4,5-trimethoxyphenylbromide, BuLi, THF; (e) 2 N HC1
in 1,4-dioxane.
[0030] Figure 7 depicts a synthetic scheme for the preparation of Aryl-
Benzoyl-Imidazole (ABI)
compounds of this invention. Reagents and conditions: (a) t-BuOH, 12,
ethylenediamine, K2CO3,
. 15 reflux; (b) Ph! (0Ac)2, K2CO3, DMSO; (c) DBU, CBrC13, DMF; (d) NaH,
PhS02C1, THF, 0 C -
RT; (e) t-BuLi, substituted benzoyl chloride, THF, -78 C; (f) Bu4NF, THF, RT.
[0031] Figure 8 depicts a synthetic scheme for the preparation of Aryl-
Benzoyl-Irnidazole (ABI)
compounds of this invention. Reagents and conditions: (a) NH4OH, oxalaldehyde,
ethanol, RT; (b)
NaH, PhS02C1, TI-IF, 0 C - RT; (c) t-BuLi, substituted benzoyl chloride, THF,
-78 C; (d) Bu4NF,
THF, RT; (e) BBr3, CH2C12; c-HC1, AcOH, reflux.
[0032] Figure 9 depicts a synthetic scheme for the preparation of Aryl-
Benzoyl-Imidazole (ABI)
compounds of this invention. Reagents and conditions: (a) NaH, substituted
benzoyl chloride, THF.
[0033] Figure 10 depicts the synthetic scheme of compounds 12dc, 12fc,
12daa, 12dab, 12cba.
(a) AlC13, THF, reflux; (b) NaH,.CH3I for 12dab and 12cba and BnBr for 12daa,
THF, reflux.
Figure 11 depicts the synthetic scheme of compounds llgaa, 121a. (a) NHIOH,
ethanol, glyoxal,
RT; (b) NaH, substituted PhS02C1, 11-1F, 0 C - RT; (c) t-BuLi (1.7 M in
pentane), substituted
benzoyl chloride, THF, -78 C; (d) Bu4NF, RT.
CA 3030689 2019-01-18
[0034] Figure 12 depicts the synthetic scheme of compound 15xaa and 12xa. (a)
1. KOH,
ethanol; 2. PhS02C1, acetone; (b) N1-L4011, glyoxal, ethanol, RT; (c) NaH,
PhS026, THF, 0 C ¨
RT; (d) t-BuLi (1.7 M in pentane), benzoyl chloride, THF, -78 "V; (e) Na0H,
ethanol, 1120, reflux.
[0035] Figure 13 depicts synthetic scheme of 17ya. (a) 1. KOH, ethanol,
2. PhS02C1, acetone,
RT; (b) NH4OH, glyoxal, ethanol, RT; (c) Nail, PhS02C1, THE, 0 C ¨ RT; (d) t-
BuLi (1.7 M in
pentane), benzoyl chloride, THF, -78 C; (e) Na0H, ethanol, H20, reflux.
[0036] Figure 14 depicts synthetic scheme of 12fa. (a) NH4OH,
oxalaldehyde, ethanol, RT; (b)
NaH, PhS02C1, THF, 0 C ¨ RT; (c) t-BuLi, 3,4,5-trimethoxybenzoyl chloride,
THF, -78 C; (d)
Bu4NF, THF, RT.
[0037] Figure 15 depicts a synthetic scheme of compound 55.
[0038] Figure 16 depicts a synthetic scheme of isoquinoline and quinoline
based compounds.
Figure 16A depicts the synthetic scheme of isoquinoline derivatives. Reagents
and conditions: a)
arylboronic acid (1 equiv.), Pd(PPh3)4 (0.01 equiv.), K2CO3, H20, DMF, 5 h; b)
arylboronic acid
(2.4 equiv.), Pd(PPh3)4 (0.04 equiv.), K2CO3; H20, DMF, 16 h; c) arylboronic
acid (1.2 equiv.),
.. Pd(PPh3)4. (0.04 equiv.), K2CO3, H20, DMF, 16 h. Figure 16B depicts the
synthetic scheme of
compounds 41 and 44. Reagents and conditions: a) p-fluorobenzenesulfonyl
chloride, pyridine,
Pyridine, 80 C, 3 h; h) 5-indoleboronic acid (1.2 equiv.), Pd(PPh3)4 (0.02
equiv.), K2CO3, H20,
DMF, 16 h. Figure 16C depicts the synthetic scheme of isoquinoline derivative
6d. Figure 16100
depicts the synthetic scheme of isoquinoline derivative 6c. Figure 16E depicts
the synthetic scheme
of isoquinoline derivative 6b.
[0039] Figure 17 depicts a standard solubility curve for ABI compound
12ga (dissolved in
acetonitrile). X-axis is the amount of compound and y-axis is the raiz peak
area.
[0040[ Figure 18 depicts the measured aqueous solubility for anti-tubulin
compounds lh, k,
66a, 2r-HCI, 5a, and 5c.
[0041] Figure 19 depicts the structures of colchicine-binding site tubulin
inhibitors.
[0042] Figure 20 depicts the ability of anti-tubulin compounds lh, lc,
2j, 66a and 5a to inhibit
tubulin polymerization in vitro (Figure 20a) and Sc (Figure 20b), and the 5Hc
binding to colchicine
site (Figure 20c).
11
CA 3030689 2019-01-18
[0043] Figure 21 depicts dose-response curves of 2-aryl-4-benzoyl-
imidazole compounds
CAB Is) compared with other anticancer drugs and compounds on multidrug
resistant melanoma cell
line (MDR cell) and the matched sensitive parent cell line (Normal Melanoma
cell). The large
distance between the two curves for paclitaxel, vinblastine, and colchicine
indicates that they were
substrates for P-glycoprotein (P-gp). The overlapping two curves of each ABI
compound indicate
that the ABI compounds were not substrates for P-gp and overcame multidrug
resistance.
[0044] Figure 22 presents the effect of ABI compounds on tubulin
polymerization in vitro.
Tubulin (0.4 mg/assay) was exposed to 10 uM ABI compounds (vehicle control, 5%
DMSO).
Absorbance at 340 nm was monitored at 37 C every minute for 15 min and
demonstrated that ABI
compounds 12da, lalb, and 12cb inhibited tubulin polymerization in vitro.
[0045] Figure 23 depicts B16-F1 melanoma colony formation assay in soft agar
which showed
that ABE compounds inhibited colony formation in a concentration-dependent
manner. Figure 23A
depicts representative pictures of control and each tested compound (12cb,
12da, and 12113) at 100
riM. The diameter of each well was 35 mm. Figure 23B depicts a quantified
representation of assay
results for each tested compound (12cb, 12da, and 12f13). P value was
calculated comparing with
control using Student's t test by GraphPad Prism software. Columns, means of
three replicates; bars,
SD.
[0046] Figure 24 depicts in viva study of ABI compounds. Figure 24A depicts
the in vivo
activity of 12cb against B16-F1 melanoma tumors in C57/BL mice. Figure 24B
depicts the in vivo
activity of 12113 against B16-FI melanoma in C57BU6 mice and SHO nude mice.
Results showed
that 12f1) inhibited melanoma tumor growth in a dose-dependent manner. C57BU6
mice bearing
B16-F1 melanoma allograft (n=5 per group). Each mouse received 0.5x106 cells
by s,c. injection into
the flank. 30 uL i.p. daily treatments were started when tumor size reached
¨100 mm3. Figure 24C
depicts the in vivo activity of 12Th against an A375 human melanoma xenograft.
SHO nude mice
bearing an A375 human melanoma xenograft (n=5 per group). Each mouse received
2.5x106cells by
s.c. injection into the flank. 30 pL i.p. daily treatments were started when
the tumor size reached
¨150 mm3. Control, vehicle solution only; points, means; bars, SD. DTIC, (5-
(3,3,-dimethy1-1-
=
triazeny1)-imidazole-4-carboxamide, dacarbazine.
[0047] Figure 25 depicts a competitive colchicine binding assay. Figure
25A depicts a [3H]-
colchicine competition-binding scintillation proximity assay which showed that
12th competitively
12
CA 3030689 2019-01-18
bound to tubulin colchicine binding site. Figure 25B depicts representative
graphs of cell cycle
analysis using flow cytometry which showed that ABI compounds (examples shown
for 12da and
12fb) arrested A375 cells in the G2/M phase after 24 h incubation. The effect
and potency were
similar to those of colchicine. Figure 25C shows quantified graphic depictions
of cell cycle analysis.
All tested compounds (examples shown for 12cb, 12da, and 12fb) arrested A375
cells in the G2/M
phase in a dose-dependent manner. ABI 12da showed greater potency than did
colchicine. Figure
25D depicts a cell cycle analysis using flow cytometry of A375 cells after
being incubated with
12cb, 12da, and 12th at different concentrations for 24 h. Colchicine arrested
most cells in the
G2/M phase starting from 50 nM. 12cb, 12da, and 12th also arrested most cells
in the G2/M phase
starting from 200, 50, and 200 nM respectively.
[0048] Figure 26 depicts the effect of 17ya and 55 on tubulin
polymerization. Compounds 17ya
and 55 bind to colchicine-binding site on tubulin, and inhibit tubulin
polymerization. Figure 26A,
competitive mass binding. Tubulin (1 mg,/mL) and colchicine (1.2 p.M) were
incubated with various
concentrations of podophylltoxin, vinblastine, compounds 17ya, and 55. N -= 3;
mean SD.
Podophylltoxin and vinblastine were used as positive and negative controls,
respectively. Figure
2613, effect on tubulin polymerization. Tubulin (0.4 mg) was exposed to test
compounds (5 tiM).
Colchicine was used as positive control. Figure 26C and 26D, ability of 17ya
and 55 to enhance
cytoplasmic DNA-Histone complex formation (apoptosis) at 24 h in PC-3 (C) and
PC-3/TxR (D)
cells (N =3); mean SD. Docetaxel was used as positive control.
[0049] Figure 27 depicts in vivo anticancer efficacy. Figure 27A, Nude mice
bearing PC-3
tumors were treated with docetaxel (i.v., 10 or 20 mg/kg) on day 1 and 9. (N =
5-6). 'Bars, SE.
Figure 27B, Nude mice bearing PC-3/TxR tumors were treated with docetaxel
(i.v., 10 or 20 mg/kg)
on day 1 and 9, compound 17ya treatments (p.o., 6.7 mg,/kg) once daily, five
days a week. (N = 4-5).
Bars, SE. Figure 27C, Nude mice bearing PC-3/TxR tumors were treated with
compound 17ya (PO,
3.3 mg/kg) twice a day for four days in the first week, and then dosed once a
day, five days a week
for weeks 2-4 (N = 7), with compound 55 treatments (p.o., 10 or 30 mg/kg)
twice a day, five days a
week for four weeks (N = 7). Bars, SE. Figure 271), Nude mice bearing PC-3/TxR
tumors were
treated with compound 17ya (PO, 10 mg/kg) three times a week for four weeks (N
= 5). Bars, SE.
[0050] Figure 28 depicts that compounds 111, 2k, and 21 inhibit tubulin
polymerization via
.. binding to the colchicine binding site on tubulin. (Figure 28A) Structures
of lh (-H), 2k (-F), and 21
(¨OH). (Figure 28B) Effect of the compounds on tubulin polymerization. Tubulin
(0.4 mg) was
13
CA 3030689 2019-01-18
exposed to compounds 1h, 2k, and 21(10 pM). Absorbance at 340 am was monitored
every min for
15 min. (Figure 28C) Ability of lh to compete for colchicine, vinblastine and
paclitaxel binding
sites on tubulin using mass spectrometry competitive binding assay (n = 3);
bars, SD.
[0051] Figure 29
depicts that compounds lh, 2k and 21 arrested cells into G2/M phase and
induced apoptosis. (Figure 29A) Representative graphs of cell cycle analysis
after compounds
treatment for 24 h on PC-3 and A375 cells. (Figure 29B) The changes in G2/M
proportion induced
by lh, 2k, and 21 in PC-3 and A375 cells after 24 h treatment. (Figure 29C)
Ability of lh, 2k, and
21 to enhance cytoplasmic DNA-Histone complex formation in 24 h (n = 3); bars,
SD. Colchicine
and vinblastine were used as positive controls.
[0052] Figure 30
depicts pharmacokinetic studies of lh, 2k and 21 administered i.p. in mice and
rats. (Figure 30A) Concentration-time curve of SMART compounds in ICR mice (n
= 3); bars, SD.
SMART compounds were administrated 15 mg/kg i.v. by tail vein injection.
(Figure 30B)
Concentration-time curve of lh and 2k in SD rats (n = 4); bars, SD. Spague-
Dawley rats were dosed
2.5 mg/kg i.v. with the formulation DMSO/PEG300 (1/4).
[0053] Figure 31
presents in vivo anti-cancer efficacy (administered i.p.) and neurotoxicity of
SMART compounds in mice. (Figure 31A) SMART compounds efficacy for PC-3
prostate tumor
xenografted on nude mice (n = 6-8). (Figure 31B) Vinblastine efficacy for PC-3
prostate tumor
xenografted on nude mice (n = 8). This served as the positive control. (Figure
31C) /n viv6 efficacy
of lh and 2k in nude mice bearing A375 melanoma xenografts (n = 10). Nude mice
were inoculated
with 2.5 x 106 PC-3 or A375 cells and dosed i.p. daily (SMART compounds) and
q2d (vinblastine)
after tumor formation (150-200 mm3). Each point represents mean tumor volume
for animals in each
group. (Figure 31D) In vivo neurotoxicity (rotarod test) of lh in ICR mice (n
= 7 or 8). lh (5 and 15
mg/kg), vinblastine (0.5 mg/kg) and vehicle were given i.p. daily, and
vinblastine was used as the
positive control. The dosing was stopped on day 3L *, p < 0.05. Bars, SE.
[0054] Figure 32 depicts
molecular modeling of ABI compounds that target tubulin in the
colchicine binding site. Figures 32A and 32B depict molecular modeling of
compound 12cb and
11cb, respectively.
[0055] Figure 33
depicts microscopic images of inununofiuorescence-labeled microtubules in
WM-164 melanoma cells, which showed microtubule modality was dramatically
changed after
14
CA 3030689 2019-01-18
compound treatment for 18 h. This provides visual proof that AM compounds
target tubulin and
disrupt functional microtubule formation.
[0056] Figure 34 depicts the efficacy and tolerability of 6b and 6c in
xenograft models after i.p.
injection. Figure 34A. PC-3 xenografts were treated with vehicle (qd), 6b (40
mg/kg, qd), or 6c (40
mg/kg, qd) for 3 weeks. Dosing vehicles were composed of 20% Captex200 in
Tween80. The tumor
volumes (mm3) were plotted against time and are the means SD from eight
animals. The tumor
volumes were shown in left panel and body weights were shown in right panel.
Figure 34B. The
liver size (g) of each nude mouse was measured after 3 weeks treatment. Figure
34C. The number
of white blood cells was counted in whole blood collected from animal after 3
weeks treatment.
[0057] Figure 35- Compound 17ya showed potent endothelial cell growth
inhibition. Cell
growth inhibition of doxorubicin (Figure 35A) and compound 17ya (Figure 35B)
was
investigated in several cell lines by SRB study. The definitions HUVEC-active
and HUVEC-
inactive represent growth factor¨supplemented and growth factor¨deprived
endothelial cell
cultures, respectively.
[0058] Figure 36- Disruption of preformed capillary by 17ya. HUVEC cells
loaded on
Matrigel were allowed to make tube for 16 h and the test compound was treated
to the preformed
tubes. The number of tubes (A, B, and C) and nodes (D, E, and F) were counted
up to 25 h after
drug treatment. Panels A and D are conditions in the presence of CA4, panels B
and E are
conditions in the presence of doxorubicin and panels C and F are conditions in
the presence of
17 ya.
[0059] Figure 37- Inhibition of the endothelial capillary formation and
disruption of
preformed capillaries. Inhibition of capillary formation (III) and disruption
Of preformed
capillary (0) were compared in vitro study using HUVEC cells after 15 h CA4 (A
and D), DOX
(B and E), and 17ya (C and F) treatment. Arrow shows the IC50 value of each
compound in ,
HUVEC cell growth inhibition.
[0060] Figure 38- 17ya and 55 increased the permeability of endothelial cell
monolayers.
Confluent HUN/EC monolayers were exposed to test compound. The leakage of ETC-
conjugated
dextran through the monolayer was assessed by relative fluorescence
measurements at X = 485
nm excitation and X. = 530 nm emission in a receiver to determine changes in
monolayer
permeability following exposure.
CA 3030689 2019-01-18
[0061] It will be appreciated that for simplicity and clarity of
illustration, elements shown in the
figures have not necessarily been drawn to scale. For example, the dimensions
of some of the
elements may be exaggerated relative to other elements for clarity. Further,
where considered
appropriate, reference numerals may be repeated among the figures to indicate
corresponding or
analogous elements.
DETAILED DESCRIPTION OF THE INVENTION
[0062] In one embodiment, this invention is directed to a compound of formula
(I)
0 ____________________________
,0
(I)
wherein
A and C are each independently substituted or unsubstituted single-, fused- or
multiple-ring aryl
or (hetero)cyclic ring systems; substituted or unsubstituted, saturated or
unsaturated N-
IS heterocycles; substituted or unsubstituted; saturated or unsaturated S-
heterocycles; substituted or
unsubstituted, saturated or unsaturated 0-heterocycles; substituted or
unsubstituted, saturated or
unsaturated cyclic hydrocarbons; or substituted or unsubstituted, saturated or
unsaturated mixed
heterocycles;
20 B is
(R10)1 (R10)1
Ri (I:tic=)1
J.J(II
N \
,
RI, (thiazole) H
(thiazole), (thiazolidine),
(R101
R
0 (R10)1 ( ioh 0-1-\
1'14 ,Ri
try' (oxazole), (ox azol ine), H (oxazolidine),
(R10)1 (Rio), (R101 R11
3-0 N HN
I N
is, (benzene), R11 (benzene) (pyrimidine),
(imidazole),
16
=
CA 3030689 2019-01-18
(R10)1
(ROI (R10)1
-N
Rii S /71 1 c-741 4==<
11
(pyridine), = / (furan), (thiophene), (R10/1
R11 (isoxazole),
(R10)1 tr"
Lyl-NI.Ri YN.,), (R10)1 (R)I
\-N R11 --NN \Fll
(piperidine), (R10)1 Rut (pyrazole), N (indole), or
k.,N1
(isoquinoline);
R10 and R11 are independently hydrogen, 0-alkyl, 0-haloalkyl, .F, Cl, Br, I,
haloalkyl, CF3, CN, -
CH2CN, NH2, hydroxyl, -(CH2)1NHCH3, -(CH2)1NFI2, -(CH2)1N(CH3)2, -0C(0)CF3, C1-
05 linear
or branched alkyl, haloalkyl, alkylamino, aminoalkyl, -OCH2Ph, -NHCO-alkyl,
COOH, -C(0)Ph,
C(0)0-alkyl, C(0)H, -C(0)NH2 or NO2;
X is a bond, NH, C1 to Cs hydrocarbon, 0, or S;
Y is a bond, -C=0, -C=S, -C=N-NH2, -C=N-OH, -CH-OH, -C=CH-CN,
-CN-CN, -CH=CH-, -C=C(CH3)2, -C=N-0Me, -(C=0)-NH, -NH-(C=0), -(C=0)-0, -0-
(C=0),
-(CH2)1-s-(C=0), (C=0)-(0-12)i-5, -(S02)-NH-, -NH-(S02)-, SO2, SO or S;
wherein said A and C rings are optionally substituted by 1-5 substituents
which are independently
0-alkyl, 0-haloalkyl, F, CI, Br, I, haloalkyl, CF3, CN, -CH2CN, NH2, hydroxyl,
-(CH2)1NHCH3, -
(CH2)1NH2, -(CH2)1N(CH3)2, -0C(0)CF3, C1-C3 linear or branched alkyl,
haloalkyl, alkylamino,
aminoalkyl, -OCH2Ph, -NHCO-alkyl, COOH, -C(0)Ph, C(0)0-alkyl, C(0)H, -C(0)NH2
or NO2;
i is an integer between 0-5;
in an integer between 1-2;
wherein
if B is a benzene ring, a thiophene ring, a furan ring or an indole ring then
X is not a bond or
CH2, and A is not indole;
if B is indole then Xis not 0; and
or its pharmaceutically acceptable salt, hydrate, polymorph, metabolite,
tautomer or isomer.
[0063] In one embodiment, if B of formula I is a thiazole ring then X is not a
bond.
[0064] In one embodiment, A in compound of Formula I is indolyl. In another
embodiment A is
2-indolyl. In another embodiment A is phenyl. In another embodiment A is
pyridyl. In another
17
CA 3030689 2019-01-18
embodiment A is naphthyl. In another embodiment A is isoquinoline. In another
embodiment, C
in compound of Formula I is indolyl. In another embodiment C is 2-indolyl. In
another
embodiment C is 5-indolyl. In another embodiment, B in compound of Formula I
is thiazole. In
another embodiment, B in compound of Formula I is thiazole; Y is CO and X is a
bond. Non
limiting examples of compound of formula I are selected from: (2-(1H-Indo1-2-
yl)thiazol-4-
y1)(1H-indol-2-yOmethanone (8) and (2-0 H-indo1-2-yOthiazol-4-y1)(1H-indol-5-
yOmethanone
(21).
[0065] In one embodiment, this invention is directed to a compound of formula
(la)
R3
B
R2
(R1)m
(Ia)
wherein
A is substituted or unsubstituted single-, fused- or multiple-ring, aryl or
(hetero)cyclic ring
systems; substituted or unsubstituted, saturated or unsaturated N-
heterocycles; substituted or
unsubstituted, saturated or unsaturated S-heterocycles; substituted or
unsubstituted, saturated or
unsaturated 0-heterocycles; substituted or unsubstituted, saturated or
unsaturated cyclic
hydrocarbons; or substituted or unsubstituted, saturated or unsaturated mixed
heterocycles;
B is
s (Rio)! (Ria
aR11 1-4 ir(R10). S-ii R
(thiazole), R11 (thiazole), H (thiazolidine),
(R101
(Ria
0.7,1(Riah 0-I-I a
j 113_Rii .xii
N
tcRii
(oxazole), N (ox azoline), H (oxazolidine),
18
CA 3030689 2019-01-18
(R10)1 (F210)1 (R10)1
HN
(benzene), R11 (benzene)
(pyrimidine), (imidazole),
(R10)1 (Rio); (Rio),
Sin
N=c
R11 c ii (R10/1 \-0'
(pyridine), )r(furan), (thiophene), -11 (isoxazole),
(R10)1 s=Pr'
7-1-Ni. RI i (ROI LP-NH (Rio), ?ii
) t Val
\--N 1211 o
(piperidine), 1.=10/1 R11 (pyrazole), N (indole), or
R11
/IX/1
vN
(isoquinoline);
Ith R2 and R3 are independently hydrogen, 0-alkyl, 0-haloalkyl, F, Cl, Br, I,
haloalkyl, CF3, CN,
-CH2CN, NH2, hydroxyl, -(CH2)1NHCH3, -(CH2)1NFI2, -(CH2)N(CH3)2, -0C(0)CF3, C1-
05 linear
or branched alkyl, haloalkyl, alkylamino, aminoalkyl, -OCH2Ph, -NHCO-alkyl,
COOH, -C(0)Ph,
C(0)0-alkyl, C(0)H, -C(0)NH2 or NO2;
Ric, and Rini are independently hydrogen, 0-alkyl, 0-haloalkyl, F, Cl, Br, I,
haloalkyl, CF3, CN, -
CH2CN, N1-12, hydroxyl, -(CH2)1NHCH3, -(CH2)1NI12, -(CH2)1N(CH3)2, -0C(0)CF3,
CI-Cs linear
or branched alkyl, haloalkyl, alkylamino, aminoalkyl, -OCH2Ph, -NHCO-alkyl,
COOH, -C(0)Ph,
C(0)0-alkyl, C(0)H, -C(0)NH2 or NO2;
X is a bond, NH, Ci to C5 hydrocarbon, 0, or S;
Y is a bond, -0=0, -C=S, -C=N-NH2, -C=N-OH, -CH-OH, -C=CH-CN,
-C=N-CN, -CH=CH-, -C=C(CH3)2, -C=N-01\4. e, -(C=0)-NH, -NH-(0=0), -(0=0)-0, -0-
(C=0),
-(CH2)1-s-(C=0), (C=0)-(CH2)1-5, -(S02)-NH-, -NH-(S02)-, SO2, SO or S;
wherein said A ring is optionally substituted by 1-5 substituents which are
independently 0-alkyl,
0-haloalkyl, F, Cl, Br, I, haloalkyl, CF3, CN, -CH2CN, NI-12, hydroxyl, -
(CH2)1NHCH3, -
(CH2)1NH2, -(CH2)iN(CH3)2, -0C(0)CF3, Ci-05 linear or branched alkyl,
haloalkyl, alkylamino,
aminoalkyl, -OCH2Ph, COOH, -
C(0)Ph, C(0)0-alkyl, C(0)H, -C(0)NH2 or NO2;
i is an integer between 0-5;
I is an integer between 1-2;
m is an integer between 1-3;
19
CA 3030689 2019-01-18
wherein
if B is a benzene ring, a thiophene ring, a furan ring or an indole ring then
X is not a bond or CH2
and A is not indole;
if B is indole then X is not 0;
or its pharmaceutically acceptable salt, hydrate, polymorph, metabolite,
tautomer or isomer.
[0066] In one embodiment, if B of formula la is a thiazole ring then X is not
a bond.
[0067] In one embodiment, this invention is directed to a compound of formula
(II):
R6
R5
(R4)n
R2 R3
II
wherein
B is
S/(Rio)i
, (Rio)1
1_4 4L-R11 ______________ (R10)1 S-11 Rii
(thiazole), R11 (thiazole), (thiazolidine);
(RioN
(R10)1
0-1-1
::117.711
N (oxazole), (oxazoline), H (oxazolidine),
(R101 (R10)1 (R10)1HN
p.
N
¨ R1(
-"N (benzene), ¨ Ril (benzene) (pyrimidine),
(irnidazole),
CA 3030689 2019-01-18
(R10)1 (Riah (Rio), O.N
iRiis S,õ
14--K0'31N; (furan), Rti (R10/1 '
(pyridine), (thiophene), R11
(isoxazole),
(Rio)!
_</N-NH Rit
A'Areci ICEI3541/4 Ril
t.µ10,1
(piperidine), R11 (pyrazole), NJ (indole), or
1-Cta'
(isoquinoline);
RI, R2, 113, Ra, R5 and R6 are independently hydrogen, 0-alkyl, 0-haloalkyl,
F, Cl, Br, I,
haloalkyl, CF3, CN, -CH2CN, NH2, hydroxyl, -(CH2),NHCH3, -(CH2)1NH2, -
(CH2)1N(CH3)2, -
OC(0)CF3, C1-05 linear or branched alkyl, haloalkyl, alkylatnino, aminoalkyl, -
OCH2Ph, -
NHCO-alkyl, COOH, -C(0)Ph, C(0)0-alkyl, C(0)H, -C(0)NH2 or NO2;
R10 and R11 are independently hydrogen, 0-alkyl, 0-haloalkylõ F, Cl, Br, 1,
haloalkyl, CF3, CN,
CH2CN, NH2, hydroxyl, -(CH2)1NHCH3, -(CH2)1NH2, -(CH2)1N(CH3)2, -0C(0)CF3, C1-
05 linear
or branched alkyl, haloalkyl, alkylarnino, aminoalkyl, -OCH2Ph, -NICO-alkyl,
COOH, -C(0)Ph,
C(0)0-alkyl, C(0)H, -C(0)NH2 or NO2;
X is a bond, NH, CI to C5 hydrocarbon, 0, or S;
Y is a bond, -C=0, -C=S, -C=N-NH2, -C=N-OH, -CH-OH, -C=CH-CN,
-C=N-CN, -CH=CH-, C=C(CH3)2, -C=N-0Me, -(C=0)-NH, -NH-(C=0), -(C=0)-0, -0-
(C=0),
-(CE12)i-5-(C=0), (C=0)-(CH2)1-5, -(S02)-NH-, -NH-(S02)-, SO2, SO or S;
i is an integer between 0-5;
1 is an integer between 1-2;
n is an integer between 1-3; and =
m is an integer between 1-3;
wherein
if B is indole then X is not 0;
or its pharmaceutically acceptable salt, hydrate, polymorph, metabolite,
tautomer or isomer.
=
[0068] In one embodiment, if B of formula II is a thiazole ring then X is not
a bond.
[0069] In one embodiment, this invention is directed to a compound of formula
(III)
21
CA 3030689 2019-01-18
R6
R5 .
X B .
y
(R4)
Me0 (110 OMe
OMe
= (III)
wherein
B is
c(Ria (R10)1
S
-+A DZ
i---4
Ns,s''Rii NN N e
e (thiazole), R11 (thiazole), I-1 (thiazolidine),
(R101
(Rich
cy,(Ria 0T-R11
11 / (oxazole), (oxazoline), H (oxazolidine),
(R10)1 (R10)1 (R10)1
N.1
HN -)----A
¨R ii i-C1-\-1 1-{'.., N
----..õ 0 ...--,4-
..õ \
(benzene), '''R11 (benzene) -,-^' (pyrimidine), s."--
(imidazole),
(Rio)i (R10)1 (R10)1 P-N
I--;,, =-= , S i,
N=--
-,..,=, (pyridine), 1 s'ss5 (fun), se (thiophene),
1411 (isoxazole),
(R10>1. ri-5_,
R11 -(R10)1
...., ,,,
1 ,,-
"-N Rõ `--N (111-4)(iN:41/
(pyrazole),(R1:7)I '''. ) RiN; (indole), Or
' (piperidine),
-S-c......1
.
>=^' (isoqUinOline);
=
22
CA 3030689 2019-01-18
R4, Rs and R6 are independently hydrogen, 0-alkyl, 0-haloalkyl, F. Cl, Br, I.
haloalkyl, CF3, CN,
-CH2CN, NH2, hydroxyl, -(CH2)1NHCH3, -(CH2);NH2, -(CH2)N(CH3)2, -0C(0)CP3, C1-
05 linear
or branched alkyl, haloalkyl, alkylamino, arninoalkyl, -OCH2Ph, -NHCO-alkyl,
COOH, -C(0)Ph,
C(0)0-alkyl, C(0)H, -C(0)NH2 or NO2; and
R10 and R11 are independently hydrogen, 0-alkyl, 0-haloalkyl, F, Cl, Br, 1,
haloalkyl, CF3, CN, -
CH2CN, NH2, hydroxyl, -(CH2)1NHCH3, -(CH2)1NH2, -(CH2)N(CH3)2, -0C(0)CF3, C1-
05 linear
.
or branched alkyl, haloalkyl, alkylamino, aminoalkyl, -0CH2Ph, -NHCO-alkyl,
COOH, -C(0)Ph,
C(0)0-alkyl, C(0)H, -C(0)NH2 or NO2;
X is a bond, NH, C1 to C5 hydrocarbon, 0, or S;
Y is a bond, -C=0, -C=S, -C=N-NH2, -C=N-OH, -CH-OH, -C=CH-CN,
-CN-CN, -CH=CH-, C=C(CH3)2, -C=N-0Me, -(C=0)--NH, -NH-(C=0), -(C=0)-0, -0-
(C=0),
-(CH2)1.5-(C=0), (C=0)-(CH2)1-5, -(S02)-NH-, -NH-(S02)-, SO2, SO or S;
i is an integer between 0-5;
I is an integer between 1-2; and
n is an integer between 1-3;
wherein
if B is indole then X is not 0;
or its pharmaceutically acceptable salt, hydrate, polymorph, metabolite,
tautomer or isomer.
[0070] In one embodiment, if B of formula III is a thiazole ring then X is not
a bond.
[0071] In one embodiment, this invention is directed to a compound of formula
(IV)
'ndole = R2 =
(IV)
wherein ring A is an indolyl;
B is
23
CA 3030689 2019-01-18
s y1:110)1 (R10)1
S.-x.)4z
S-I-N R
;R11 F-4 ir(Rio)1 ws. -:-L ii
N"--`,,s5
c' (thiazole), R11 (thiazole), H (thiazolidine),
(Ria
0(F110)1
---- +-1211
Y(N),111
- r (oxazole), \ N Or (oxazoline), H
(oxazolidine),
(R10)1 (Ria (R10)1
N1-,) FiN1"'s4r--A
1._ )sIs(R10)1
"-(- p ..11 -<.,11--1
(benzene), R11 (benzene) -,.-= (pyrimidine), "'",-
(imidazole),
(R10)1 (R10)1 (Ria
(Ria N-1 'sr'
=N"' (pyridine), (furan), / (thiophene), R11
(isoxazole),
(RNA
/I- \1Rii >-,,...,-(Rioh
N-NH (R10)1 Ril
r\--k--1-:\
AA-ki. µL _ 1
\-N R11 \--N (R10)1
. ;,, , -%,"' (piperidine), R-11' (pyrazole), 1-
-.%- -1µ1 (indole), or
(Rioh \ , ylli,
(isoquinoline);
R1 and R2 are independently hydrogen, 0-alkyl, 0-haloalkyl, F, Cl, Br, I,
haloalkyl, CF3, CN, - =
CH2CN, NH2, hydroxyl, -(CH2);NHCF13, -(CH2)1NF12, -(CH2)1N(0-13)2, -0C(0)CF3,
CI-Cs linear
or branched alkyl, haloalkyl, alkylamino, aminoalkyl, -OCH2Ph, -NHCO-alkyl,
COOH, -C(0)Ph,
C(0)0-alkyl, C(0)H, -C(0)N112 or NO2; .
Rio and Rii are independently hydrogen, 0-alkyl, 0-haloalkyl, F, CI, Br, I,
haloalkyl, CF3, CN, -
CH2CN, NH2, hydroxyl, -(CH2),NECH3, -(CH2)1NH2, -(CH2)N(CH3)2, -0C(0)CF3, C1-
05 linear
or branched alkyl, haloalkyl, alkylamino, aminoalkyl, -OCH2Ph, -NHCO-alkyl,
COOH, -C(0)Ph,
C(0)0-alkyl, C(0)H, -C(0)NH2 or NO2;
X is a bondõ NH, C1 to C5 hydrocarbon, 0, or S;
Y is a bond, C=0, -C=S, -C=N-NH2, -C=N-OH, -CH-OH, -C=CH-CN, .
-C=N-CN, -CH=CH-, C=C(CH3)2, -C=N-0Me, -(C=0)-NH, -NU-(C=O), -(C=0)-0, -0-
(C=0),
-(CH2)1_5-(C=0), (C=0)-(CF12)1-5, -(S02)-NH-, -NI-1-(S02)-, SO2, SO or S;
24
CA 3030689 2019-01-18
wherein said A is optionally substituted by 0-alkyl, 0-haloalkyl, F, Cl, Br,
1, haloalkyl, CF3, CN,
-CH2CN, NH2, hydroxyl, -(CH2)1NHCH3, -(CH2)1NH2, -(CH2)1N(CH3)2, -0C(0)CF3, CI-
05 linear
or branched alkyl, haloalkyl, alkylainino, amincialkyl, -OCH2Ph, -NHCO-alkyl,
C001-1, -C(0)Ph,
C(0)0-alkyl, C(0)H, -C(0)NH2 or NO2; and
i is an integer between 0-5;
1 is an integer between 1-2; and
m is an integer between 1-4;
wherein
if B is a benzene ring, a thiophene ring, a furan ring or an indole ring then
X is not a bond or
CH2;
or its pharmaceutically acceptable salt, hydrate, polymorph, metabolite,
tautomer or isomer.
[0072] In one embodiment, if B of formula IV is a thiazole ring then X is not
a bond.
[0073] In another embodiment, the indolyl of ring A of formula IV is attached
to one of its 1-7
positions to X or direct to B if X is a bond (i.e nothing).
[0074] In one embodiment, this invention is directed to a compound of formula
IV(a)
HN X¨ TOA
(R4)11
R2 oil
(R
IV(a) _
B is
CA 3030689 2019-01-18
stc1R10)1
, S...1-' (R101
N
-4 -Ril N''
-.-4 !--r-(R0)1 R
--µ,55
N i
c' (thiazole), Rii (thiazole), 11 (thiazolidine),
(RIA (R10)1
cyi(Rio)i
0-1-\ Rii
-<µNI i 1)..,,,
/ (oxazole), ' N ,v (oxazoline), VI (oxazolidine),
(Rio) (Rio)i (R10)1 Rii s
Iµ N lµµ H1\1,--14
--' (benzene), R11 (benzene) ,--'
(pyrimidine), .."-i- (imidazole),
(R16)1
(Rio)i (Ri0)i _ /0
_...4-1- =-.--/R s S -N
11
4-A,
N--=c _Ã.-/ 1 i_R, k"-"A_T6 cRii
(R10),'ssr
l
Ø' (pyridine), ' 'if' (furan), r's (thiophene), RII
(isoxazole),
(Rio)! ,
LII-NIR" R11 \-N
...._<,
N-NH a Ril
/ ) A-A1-1.., (Ri µ10:V.
5 .;.,,, , -==0 (piperidine), (R10)I 13/(pyrazole), --
- N (indole), or
la
,1-' (isoquinoline);
RI, R2, RI and Rs are independently hydrogen, 0-alkyl, 0-haloalkyl, F, Cl, Br,
I, haloalkyl, CF3,
CN, -CH2CN, NH2, hydroxyl, -(CH2)1NHCH3, -(CH2)1N112, -(CH2)1N(CH3)2, -
0C(0)CF3, C1-05
linear or branched alkyl, haloalkyl, alkylamino, aminoalkyl, -OCH2Ph, -NHCO-
alkyl, COOH, -
C(0)Ph, C(0)0-alkyl, C(0)H, -C(0)H2 or NO2; and
Kw and RH are independently hydrogen, 0-alkyl, 0-haloalkyl, F, CI, Br, I,
haloalkyl, CF3, CN, -
CH2CN, NH2, hydroxyl, -(CH2);NHCH3, -(CH2);NH2, -(CH2)1N(CH3)2, -0C(0)CF3, Ci-
05 linear
or branched alkyl, haloalkyl, alkylamino, aminoalkyl, -OCH2Ph, -NHCO-alkyl,
COOH, -C(0)Ph,
C(0)0-alkyl, C(0)H, -C(0)NH2 or NO2; -
X is a bond,NH, C1 to C5 hydrocarbon, 0, or S;
Y is a bond or C=0, -C=S, -CN-NH2, -CN-OH, -CH-OH, -C=CH-CN,
-C=N-CN, -CH=CH-, C=C(CH3)2, -C=N-0Me, -(C=0)-NH, -NH-(C=0), -(C=0)-0, -0-
(C=0),
-(CH2)1_5-(C=0), (C=0)-(CH2)1-5, -(S02)-NH-, -NH-(S02)-, SO2, SO or S;
i is an integer between 0-5; .
26
CA 3030689 2019-01-18
I is an integer between 1-2;
n is an integer between 1-2; and .
in is an integer between 1-4;
wherein -
if B is a benzene ring, a thiophene ring, a furan ring or an indole ring then
X is not a bond or
CH2;
or its pharmaceutically acceptable salt, hydrate, polymorph, metabolite,
tautomer or isomer.
[0075] In one embodiment, if B of formula IVa is a thiazole ring then X is not
a bond.
[0076] In one embodiment, this invention is directed to a compound of formula
(V)
R6
Rs,B
(R4)n -
Me0 11101 OMe
OMe
(V) =
B is
(R.10)1
s/.1(R10)1
R-i Hs 10)1 S-I-\ R
N- 1i N.\
' N'y
c' (thiazole), R11 (thiazole), H
(thiazolidine),
v
(Rio)1
(R101
0.(Rio)i r-I-Nt Ri 1
0-I-A RI 1 'ii V-NiNr ,sss
N "Nar
e (oxazole), \(41µ12-/
(oxazoline), H (oxazolidine),
(R10)1 (R10)1 (R10)1 Ri 1 s
N-1-.; H N
--- - - = - -1
1,' N
¨...... R1 rs=c
.0' (benzene), R11 (benzene) Ø` (pyrimidine), 'N.
(imidazole),
(R10)1
(R101 (Rio)! 0-N
...._117-r-Rii 5 S iii --'4y,, a
N-=< _e--/-)- _R ,--kõ,-,-.-..õ711 (R10)I -pi Nse
-04 (pyridine), o-('I
r4 (thiophene), -11 (isoxazole),
27 .
CA 3030689 2019-01-18
(R10)1
N-NH (R10)1 Rit
\--N R1('-ti µ0-
(Riatv-RCliF (pyrazole), N (indole), or
, (piperidine),
(Rio,
X'11
\.N
(isoquinoline);
Ra, R5 and R6 are independently hydrogen, 0-alkyl, 0-haloalkyl, F, Cl, Br, I,
haloalkyl, CF3, CN,
-CH2CN, NH2, hydroxyl, -(CH2)1NBCF13, -(CH2)1NFI2, -(CH2)1N(CH3)2, -0C(0)CF3,
C1-05 linear
or branched alkyl, haloalkyl, alkylamino, aminoalkyl, -OCH2Ph, -NIICO-alkyl,
COOH, -C(0)Ph,
C(0)0-alkyl, C(0)H, -C(0)NH2 or NO2;
R10 and R11 are independently hydrogen, 0-alkyl, 0-haloalkyl, F, Cl, Br, I,
haloalkyl, CF3, CN, -
CH2CN, NH2, hydroxyl, -(CF12),NHCH3, -(CH2)1N1-I2, -(CH2)1N(CH3)2, -0C(0)CF3,
CI -05 linear
or branched alkyl, haloalkyl, alkylamino, aminoalkyl, -OCH2Ph, -NHCO-alkyl,
COOH, -C(0)Ph,
C(0)0-alkyl, C(0)H, -C(0)NH2 or NO2;
i is an integer between 1-5;
I is an integer between 1-2; and
n is an integer between 1-3;
or its pharmaceutically acceptable salt, hydrate, polymorph, metabolite,
tautomer or isomer,
(R10)1
õ NSki
11
[0077] In another embodiment, B of formula V is not a thiazole . In another
embodiment, B of formula V is not an oxazole. In another embodiment, B of
formula V is not an
oxazoline. In another embodiment, B of formula V is not an imidazole. In
another embodiment, B
of formula V is not a thiazole, oxazole, oxazoline or imidazole.
=
28
CA 3030689 2019-01-18
[0078] In one embodiment, this invention is directed to the following
compounds:
Formula V Compound B . 124, R5
and
R6
R5
* 0 --0 la . H
M = 40 lb
, N--
H
OMe
ome
lc e)H
N=--\
id (1.H
le N \
---S").># I-13%
0 H
PR,
. .,
If S s H
¨0, 'I
/ o
meo
OMe
Me0
lg ,-rtS__,
Q 40
N 0 H
Me0 lel
OMe
Me0
lb Sõ H
N tsis
li s 0-N H
/
29
CA 3030689 2019-01-18
lk .f=P' OMe H
0
NI c OMe
rs- HN
N. OMe
4111
11
o/
o-
35a 0 / \ 1-1
0 0
N rsji
0
0
N 0
36a 0
1-4 3 0 0
N,
0
0
NN 0
=
CA 3030689 2019-01-18
100791 In one embodiment, this invention is directed to a compound of formula
(VI)
OMe
OMe
S N OMe
(R4)fl R6
R5
(VI)
wherein
. R4, R5 and R6 are independently hydrogen, 0-alkyl, 0-haloalkyl, F, Cl,
Br, I, haloalkyl, CF3, CN,
-CH2CN, NI-I2, hydroxyl, -(CH2);NHCH3, -(CH2)1NH2, -(CH2)1N(CH3)2, -0C(0)CF3,
CI-Cs linear
or branched alkyl, haloalkyl, alkylamino, aminoalkyl, -OCH2Ph, -NHCO-alkyl,
COOH, -C(0)Ph,
C(0)0-alkyl, C(0)H, -C(0)NH2 or NO2; and
Y is a bond or C=0, -C=S, -C=N-NH2, -C=N-OH, -CH-OH,
-C=N-CN, -CH=CH-, C=C(CH3)2, -C=N-OMe, -(C=0)-NH, -NH-(C=0), ¨(C=0)-0, -0-
(C=0),
-(CH2)1-5-(C=0), (C=0)-(CH2)1-5, -(S02)-NH-, -NH-(S02)-, SO2, SO or S;
n is an integer between 1-3; and
i is an integer from 1-5;
or its pharmaceutically acceptable salt, hydrate, polymorph, metabolite,
tautomer or isomer.
[0080] In one embodiment, this invention is directed to the following
compounds:
31
CA 3030689 2019-01-18
Formula VI Compound Y ita, R5 and R6
= OM e lh -C=0
ome 2a 1C=C(CH3)2 H
2b -CH-OH
OMe
2c -C=CH-CN H
K6 (cis and trans)
R5 2d -C=N-NH2 H
(Cis and trans)
2e -C=N-OH H
(cis and trans)
= 21 -C=N-0Me H
(cis and trans)
2g -(C=0)-NH- H
2h -NH-(C=0)- H
21 nothing
2j -C=N-CN
(cis and trans) =
2k 0=0 R4=R6=H
R5=p-F
21 C=0 R4=R6=H
R5=p-OH
2m C=0 124=126=H
R5=p-CH3
2n C=0 RI=R6=H
R5=p-CH2-CN
2o C=0 124=R6=H
R5=p-N(C}13)2
2p C=0 R4=m-F;
R5=p-F;
R6=m-F;
n=1
32
CA 3030689 2019-01-18
=
2q C=0 R4=R6=H
R5=
p-CH2-(C=0)NI-12
2r C=0 R4=R6=H
R5=p-CH2NH2
2s C=0 R4=R6=H
R5=p-CH2NH-CH3
2t C=0 124=m-OMe;
R5=p-OMe;
R6=m-OMe;
n=1
2u C=0 R4=R6=1-1
R5=p-C1-12NMe2
[0081] In one embodiment, this invention, is directed to compound 3a:
S OMe
N
OMe
Me0
3a
[0082] In one embodiment, this invention is directed to compound 3b:
Me0
OMe
OMe
S N
3b
[0083] In one embodiment, this invention is directed to a compound of formula
(VII)
33
CA 3030689 2019-01-18
OMe
Y OMe
OMe
(v.)
wherein
Y is a bond or C=0, -C=N-NH2, -C=N-OH, -CH-OH, -C=CH-CN,
-C=N-CN, -CH=CH-, C=C(CH3)2, -C=N-OMe, -(C=0)-NH, -NH-(C=0), ¨(C=0)-0, -0-
(C=0),
-(CH2)1_5-(C=0), (C=0)-(CH2)1-5, -(S02)-NH-, -N11-(S02)-, SO2, SO or S;
or its pharmaceutically acceptable salt, hydrate, polymorph, metabolite,
tautomer or isomer.
[0084] In one embodiment, this invention is directed to the following
compounds:
Formula VII Compound
OMe 4a
Y 41t. OMe 4b SO2
OMe 4c SO
4d -( S02)-NH-
[0085] In one embodiment, this invention is directed to a compound of formula
(VIII)
34
CA 3030689 2019-01-18
OMe
0
\ OMe
Q N OMe
HN
(R-4)n R6
(Vin)
wherein
R4, R5 and 146 are independently hydrogen, 0-alkyl, 0-haloalkyl, F, Cl, Br, I,
haloalkyl, CF3, CN,
-CH2CN, NH2, hydroxyl, -(CH2)1NHCH3, -(CH2)1NH2, -(CH2)1N(CH3)2, -0C(0)CF3, C1-
05 linear
or branched alkyl, haloalkyl, alkylamino, aminoalkyl, -OCH2Ph, -NHCO-alkyl,
COOH, -C(0)Ph,
C(0)0-alkyl, C(0)H, -C(0)NH2 or NO2;
Q is S, 0 or NH;
i is an integer between 0-5; and
n is an integer between 1-3;
or its pharmaceutically acceptable salt, hydrate, polymorph, metabolite,
tautomer or isomer.
[0086] In one embodiment, this invention is directed to the following
compounds:
Formula VIII Compound R4 R5 146
5a
n=1
5b H p-CH3 H
n=1
Sc H p-F
n=1
. 35
CA 3030689 2019-01-18
OMe
0
OMe 5d p-C1 .S
n=1
N OMe
1
HN 5e
R6 n= I
R5
[0087] In one embodiment, this invention is directed to a compound of formula
(IX)
R5
N
A'
NH
(R-4)n
(IX)
wherein
Rs and Rs are independently hydrogen, 0-alkyl, 0-haloalkyl, F, Cl, Br, I,
haloalkyl, CF3, CN, -
CH2CN, NI12, hydroxyl, -(CH2)1NHCH3, -(CH2)2NH2,
-(CH2)1N(CH3)2, -0C(0)CF3, CI-Cs linear or branched alkyl, haloalkyl,
alkylamino, aminoalkyl, -
OCH2Ph, -NHCO-alkyl, COOK -C(0)Ph. C(0)0-alkyl, C(0)H, -(0)NH2 or NO2;
A' is halogen; substituted or unsubstituted single-, fused- or multiple-ring,
aryl or (hetero)cyclic
ring systems; substituted or unsubstituted, saturated or unsaturated N-
heterocycles; substituted or
unsubstituted, saturated or unsaturated S-heterocycles; substituted or
unsubstituted, saturated or
unsaturated 0-heterocycles; substituted or unsubstituted, saturated or
unsaturated cyclic
hydrocarbons; or substituted or unsubstituted, saturated or unsaturated mixed
heterocycles;
wherein said A' ring is optionally substituted by 1-5 substituents which are
independently 0-
alkyl, 0-haloalkyl, F, Cl, Br, I, haloalkyl, CF3, CN, -CH2CN, NH2, hydroxyl, -
(CH2)1NHCH3, -
(CH2)1NH2, -(CH21N(CH3)2, -0C(0)CF3, C1-05 linear or branched alkyl,
haloalkyl, alkylamino,
aminoalkyl, -OCH2Ph, -NHCO-alkyl, COOH, -C(0)Ph, C(0)0-alkyl, C(0)H, -C(0)NH2
or NO2;
36
CA 3030689 2019-01-18
i is an integer between 1-5; and
n is an integer between 1-3;
or its pharmaceutically acceptable salt, hydrate, polymorph, metabolite,
tautorner or isomer.
[0088] In one embodiment, a compound of Formula 1X is represented by the
structures of the
following compounds:
Formula IX Compound A' R4, R5
6a
<J5
'N 41)
H3
H3C0 OCH3
OC
A'
NH
(R4)n 6b
H
6c
6d CI
10 [0089] In another embodiment A' of formula IX is a halogen. In one
embodiment A' of formula
IX is a phenyl. In another embodiment A' of formula IX is substituted phenyl.
In another
embodiment the substitution of A' is halogen. In another embodiment the
substitution is 4-F. In
another embodiment the substitution is 3,4,5-(OCH3)3. In another embodiment,
A' of formula IX
is substituted or unsubstituted 5-indolyl. In another embodiment, A' of
formula IX is substituted
15 or unsubstituted 2-indolyl. In another embodiment, A' of formula IX is
substituted or
unsubstituted 3-indolyl. In another embodiment, compounds of formula IX are
presented in
Figure 16A.
[0090] In one embodiment, this invention is directed to a compound of formula
(IXa)
37
CA 3030689 2019-01-18
(R.On
N
A'
Rs
(IXa)
wherein
R4 and Rs are independently hydrogen, 0-alkyl, 0-haloalkyl, F, Cl, Br, I,
haloalkyl, CF3, CN, -
CH2CN, NI-12, hydroxyl, -(CH2)1NHCH3, -(CH2)iNH2,
-(CH2);N(CH3)2, -0C(0)CF3, C1-Cs linear or branched alkyl, haloalkyl,
alkylamino, aminoalkyl, -
OCH2Ph, -NHCO-alkyl, COOH, -C(0)Ph, C(0)0-alkyl, C(0)H, -(0)NH2 or NO2;
A' is halogen; substituted or unsubstituted single-, fused- or multiple-ring,
aryl or (hetero)cyclie
ring systems; substituted or unsubstituted, saturated or unsaturated N-
heterocycles; substituted or
unsubstituted, saturated or unsaturated S-heterocycles; substituted or
unsubstituted, saturated or
unsaturated 0-heterocycles; substituted or unsubstituted, saturated or
unsaturated cyclic "
hydrocarbons; or substituted or unsubstituted, saturated or unsaturated '
mixed heterocycles;
wherein said A' ring is optionally substituted by 1-5 substituents which are
independently 0-
alkyl, 0-haloalkyl, F, Cl, Br, I, haloalkyl, CF3, CN, -CH2CN, NH2, hydroxyl, -
(012)iNHCH3, -
(CH2)1NH2, -(CH2)1N(CH3)2, -0C(0)CF3, C1-05 linear or branched alkyl,
haloalkyl, alkylamino,
aminoalkyl, -OCH2Ph, -NICO-alkyl, COOH, -C(0)Ph, C(0)0-alkyl, C(0)H, -C(0)NH2
or NO2;
i is an integer between 1-5; and
n is an integer between 1-3;
or its pharmaceutically acceptable salt, hydrate, polymorph, metabolite,
tautomer or isomer.
[00911 In another embodiment A' of formula IXa is a halogen. In one embodiment
A' of
formula IXa is a phenyl. In another embodiment A' of formula IXa is
substituted phenyl. In
another embodiment the substitution of A' is halogen. In another embodiment
the substitution is
4-F. In another embodiment the substitution is 3,4,5-(OCH3)3. In another
embodiment, A' of
formula IXa is substituted or unsubstituted 5-indolyl. In another embodiment,
A' of formula IXa
is substituted or unsubstituted 2-indolyl. In another embodiment, A' of
formula IXa is substituted
or unsubstituted 3-indolyl.
38
CA 3030689 2019-01-18
[0092] In another embodiment, a compound of formula IXa is 1-chloro-7-(4-
fluorophenyl)isoquino)ine. In another embodiment, a compound of formula IXa is
7-(4-
fluoropheny1)-1-(1H-indo1-5-yOisoquinoline. In another embodiment, a compound
of formula
IXa is 7-(4-fluoropheny1)-1-(3,4,5-trimethoxyphenyl)isoquinoline. In another
embodiment, a
compound of formula IXa is 1,7-bis(4-fluorophenyl)isoquinoline (40). In
another embodiment, a
compound of formula IXa is 1,7-bis(3,4,5-trimethoxyphenyl)isoquinoline. In
another
embodiment, a compound of formula IXa is 1-(4-fluoropheny1)-7-(3,4,5-
trimethoxyphenyl)isoquinoline. In another embodiment, a compound of formula
IXa is 1-(1H-
indo1-5-y1)-7-(3,4,5-trimethoxyphenypisoquinoline. In another embodiment, a
compound of
formula IXa is 1-chloro-7-(3,4,5-trimethoxyphenyl)isoquinoline.
[0093] In one embodiment, this invention is directed to a compound represented
by the structure
of formula XI:
X--µN 0
Me0 OMe
OMe (XI)
wherein
X is a bond, NH or S:
Q is 0, NH or S; and
A is substituted or unsubstituted single-, fused- or multiple-ring aryl or
(hetero)cyclic ring
systems; substituted or unsubstituted, saturated or unsaturated N-
heterocycles; substituted or
unsubstituted, saturated or unsaturated S-heterocycles; substituted or
unsubstituted, saturated or
unsaturated 0-heterocycles; substituted or unsubstituted, saturated or
unsaturated cyclic
hydrocarbons; or substituted or unsubstituted, saturated or unsaturated mixed
heterocycles;
wherein said A ring is optionally substituted by 1-5 1-5 substituents which
are independently 0-
alkyl, 0-haloalkyl, F, Cl, Br, I, haloalkyl, CF3, CN, -CH2CN, NH2, hydroxyl, -
(CH2)1NHCH3, -
(CH2)1NH2, -(CH2),N(CH3)2, -0C(0)CF3, CI-Cs linear or branched alkyl,
haloalkyl, alkylamino,
aminoalkyl, -OCH2Ph, -NHCO-alkyl, COON, -C(0)Ph, C(0)0-alkyl, C(0)H, -C(0)N}12
or NO2;
and
39
CA 3030689 2019-01-18
i is an integer from 0-5;
or its pharmaceutically acceptable salt, hydrate, polymorph, metabolite,
tautomer or isomer.
[0094] In one embodiment if Q of Formula XI is S, then X is not a bond.
[0095] In one embodiment, A of compound of Formula XI is Ph. In another
embodiment, A of
compound of Formula XI is substituted Ph. In another embodiment, the
substitution is 4-F. In
another embodiment, the substitution is 4-Me. In another embodiment, Q of
compound of
Formula XI is S. In another embodiment, X of compound of Formula XI is NH. Non
limiting
examples of compounds of Formula XI are selected from: (2-(phenylamino)thiazol-
4-y1)(3,4,5-
trimethoxyphenypmethanone (5a), (2-(p-
tol yl amino)thi azol-4-y1)(3,4,5-
trimethoxyphenyOmethanone (5b), (2-(p-
fluorophenylamino)thiazol-4-y1)(3,4,5-
trimethoxyphenyl)methanone (5c), (2-(4-
chloropheriylamino)thiazol-4-y1)(3,4,5-
trimethoxyphenyl)methanone (5d), (2-
(phenylamino)-1H-imidazol-4-y1)(3,4,5-
trimethoxyphenyl)methanone (Sc), (2-
(phenylarnino)thiazol-4-y1)(3,4,5-
trimethoxyphenypmethanone hydrochloride salt (5Ha), (2-(p-tolylamino)thiazol-4-
y1)(3,4,5-
trimethoxyphenyl)methanone hydrochloride salt (5Hb), (2-(p-
fluorophenylamino)thiazol-4-
yl)(3,4,5-tri methox yphenyl)methanone hydrochloride salt
(5Hc), (2-(4-
chlorophenylamino)thiazol-4-y1)(3,4,5-trimethoxyphenypmethanone htdrochloride
salt (5Hd),
(2 -(phenylam ino)-1H-imidazol-4-y1)(3,4,5-trimetho xyphenypmethanone
hydrochloride salt
(5He).
= 20
[0096] In one embodiment, this invention is directed to a compound represented
by the structure
of formula XI(a):
CA 3030689 2019-01-18
OMe
0
1 OMe
HN N OMe
HN R5
(R4n
NH
XI(a)
wherein R4 and R5 are independently hydrogen, 0-alkyl, 0-haloalkyl, F, Cl, Br,
I, haloalkyl, CF3,
CN, -CH2CN, NH2, hydroxyl, -(CH2)1NHCH3, -(CH2)1NFI2, -(CH2)N(CH3)2, -
0C(0)CF3, C1-Cs
linear or branched alkyl., haloalkyl, alkylamino, aminoalkyl, -OCH2Ph, -NHCO-
alkyl, COOH, -
C(0)Ph, C(0)0-alkyl, C(0)H, -C(0)NH2 or NO2;
i is an integer from 0-5; and
n is an integer between 1-4;
or its pharmaceutically acceptable salt, hydrate, polymorph, metabolite,
tautomer or isomer.
[0097] In one embodiment, this invention is directed to a compound represented
by the structure
of formula XI(b):
=
OMe
0
OMe
HN N OMe
HN R5
(R4)n
XI(b)
wherein R4 and Rs are independently hydrogen, 0-alkyl, 0-haloalkyl, F, Cl, Br,
I, haloalkyl, CF3,
CN, -CH2CN, NH2, hydroxyl, -(CH2)1NHCH3, -(CH2)1NH2, -(0-12)1N(CH3)2, -
0C(0)CF3, C1-05
41
CA 3030689 2019-01-18
linear or branched alkyl, haloalkyl, alkylamino, aminoalkyl, -OCH2Ph, -NHCO-
alkyl, COOH, -
C(0)Ph, C(0)0-alkyl, C(0)H, -C(0)NH2 or NO2;
i is an integer from 0-5; and
n is an integer between 1-4;
or its pharmaceutically acceptable salt, hydrate, polymorph, metabolite,
tautomer or isomer.
[0098] In one embodiment, this invention is directed to a compound represented
by the structure
of formula XI(c):
OMe
0
OMe
S N OM:
HN Rs
(R4)n
NH
XI(c)
wherein R4 and R5 are independently hydrogen, 0-alkyl, 0-haloalkyl, F, Cl, Br,
I, haloalkyl, CF3,
CN, -CH2CN, NII2, hydroxyl, -(CH2),NHCH3, -(CH2)1NH2, -(CH2)1N(CH3)2, -
0C(0)CF3, C1-05
linear or branched alkyl, haloalkyl, alkylamino, aminoalkyl, -OCH2Ph, -NHCO-
alkyl, COOH, -
C(0)Ph, C(0)0-alkyl, C(0)H, -C(0)NH2 or NO2;
i is an integer from 0-5; and
n is an integer between 1-4;
or its pharmaceutically acceptable salt, hydrate, polymorph, metabolite,
tautomer or isomer.
[0099] In one embodiment, this invention is directed to a compound represented
by the structure
of formula XI(d):
42
CA 3030689 2019-01-18
=
OMe
0
OMe
S N OMe
HN
(114)n
XI(d)
wherein R4 and R5 are independently hydrogen, 0-alkyl, 0-haloalkyl, F, Cl, Br,
I, haloalkyl, CF3,
CM, -CH2CN, N112, hydroxyl, -(CH2)1NHCH3, -(CH2)iNH2, -(CH2)1N(CH3)2, -
0C(0)CF3, C1-05
linear or branched alkyl, haloalkyl, alkylamino, aminoalkyl, -OCH2Ph, -NHCO-
alkyl, COOH, -
C(0)Ph, C(0)0-alkyl, C(0)H, -C(0)NH2 or NO2;
i is an integer from 0-5; and
n is an integer between 1-4;
or its pharmaceutically acceptable salt, hydrate, polymorph, metabolite,
tautomer or isomer_
[00100] In one embodiment, this invention is directed to a compound
represented by the
structure of formula XI(e):
OMe
= 0
= OMe
_ HN N OMe
R5
HN
(R4/n XI(e)
wherein R4 and R5 are independently hydrogen, 0-alkyl, 0-haloalkyl, F, Cl, Br,
I, haloalkyl, CF3,
CN, -CH2CN, NHz, hydroxyl, -(CH2)1NHCH3, -(CH2);NH2, -(CH2)1N(CH3)2, -
0C(0)CF3, CI-05
linear or branched alkyl, haloalkyl, alkylamino, aminoalkyl, -OCH2Ph, -NHCO-
alkyl, COOH, -
C(0)Ph, C(0)0-alkyl, C(0)H, -C(0)NH2 pr NO2;
i is an integer from 0-5; and
43
CA 3030689 2019-01-18
n is an integer between 1-4;
or its pharmaceutically acceptable salt, hydrate, polymorph, metabolite,
tautomer or isomer.
[00101] In another embodiment, a compound of formula XI is represented
by the structure
of compound 55:
OMe
0
OMe
Sy N OMe
HN
NH
(55).
[00102] In another embodiment, a compound of formula XI is represented
by the structure
of compound 17ya:
OMe
0
OMe
HE
HN
(17ya).
[00103) In one embodiment, this invention provides a compound
represented by the
following structures:
compound structure
8
z
s
44
CA 3030689 2019-01-18
9
0
*
.4"
CH
3
* =
0
0
11 0 *
s
12 o
13
14 0
--F
=
CA 3030689 2019-01-18
16
0
17
0
=
F
18
o ci
Sy O¨CHa
=
19
INTrV
\¨_=/
11101
113C, 1-111
N.
P
46
CA 3030689 2019-01-18
21
q H
22
23 HC 0
r
(3--c H:
24 ?"3
0
0
0
0 -
N
47
CA 3030689 2019-01-18
26 0
- N
411 0õ, =
H2N
27
0
0
0 =
0-
28
0
N N
0
29 0
F =14
0
30 0
F *N
0
0 fa
48
CA 3030689 2019-01-18
32 NC
OMe
OH
s z N OMe
33
0
OH
O.
34 H3R
0-CI3
35 Hp-o
H3C /
- 0
CH3 p
H3C
o 0-013
tH3
49
CA 3030689 2019-01-18
40
N
41
=
0 0
42
N,
Br ,S,
0/O*
43
Br
0- I
-S
= 0
44
Nõ0
N
NH
46
. HN
NN
CA 3030689 2019-01-18
47
N
NH
48
N
NH
\ NH
49
IIIII
ANI
NH
" NH
I N
NH
H3C0 OCH3
OCH3
51 õ--
HN
0
H3C0
H3C0 OCH3
52
N N
0
H3C0
H3C0 OCH3
51
CA 3030689 2019-01-18
53
NH
0
H3C0
H3C0 OCH3
54 N-
\
0
H3C0
H3C0 ocH3
[00104] It is well understood that in structures presented in this
invention wherein the
nitrogen atom has less than 3 bonds, H atoms are present to complete the
valence of the nitrogen.
[00105] In one embodiment the A, A' and/or C groups of formula I, I(a), IV,
IX, IX(a)
and XI are independently substituted and unsubstituted furanyl, indolyl,
pyridinyl, phenyl,
biphenyl, triphenyl, diphenylmethane, adamantane-yl, fluorene-yl, and other
heterocyclic analogs
such as, e.g., pyrrolyl, pyrazolyl, imidazolyl, pyridinyl, pyrimidinyl,
pyrazinyl, pyridazinyl,
triazinyl, tetrazinyl, pyrrolizinyl, indolyl, isoquinolinyl, quinolinyl,
isoquinolinyl,
benzimidazolyl, indazolyl, quinolizinyl, cinnolinyl, quinalolinyl,
phthalazinyl, naphthyridinyl,
quinoxalinyl, oxiranyl, oxetanyl, tetrahydrofuranyl, tetrahydropyranyl,
dioxanyl, furanyl,
pyrylium, benzofuranyl, benzodioxolyl, thiranyl, thietanyl,
tetrahydrothiophene-yl, dithiolanyl,
tetrahydrothiopyranyl, thiophene-yl, thiepinyl, thianaphthenyl, oxathiolanyl,
morpholinyl,
thioxanyl, thiazolyl, isothiazolyl, thiadiazolyl, oxazolyl, isoxazolyl,
oxadiaziolyl).
[00106) In one embodiment, the A, A' and/or C groups is substituted and
unsubstituted
phenyl. In another embodiment, the A, A' and/or C groups is phenyl substituted
by Cl, F or
methyl. In one embodiment, the A, A' and/or C groups is substituted and
unsubstituted
isoquinolinyl. In one embodiment, the A, A' and/or C groups include
substituted and
unsubstituted indolyl groups; most preferably, substituted and unsubstituted 3-
indolyl and 5-
indolyl.
52
CA 3030689 2019-01-18
[00107] In one embodiment, the A, A' and/or C groups of formula I,
I(a), IV, Et, IX(a)
and XI can be substituted or unsubstituted. Thus, although the exemplary
groups recited in the
preceding paragraph are unsubstituted, it should be appreciated by those of
skill in the art that
these groups can be substituted by one or more, two or more, three or more,
and even up to five
substituents (other than hydrogen).
[00108] In one embodiment, the most preferred A, A' and/or C groups are
substituted by
3,4,5-trimethoxyphenyl. In another embodiment the A, A' and/or C groups are
substituted by
alkoxy. ln another embodiment the A, A' and/or C groups are substituted by
methoxy. In another
embodiment the A, A' and/or C groups are substituted by alkyl. In another
embodiment the A, A'
and/or C groups are substituted by methyl. In another embodiment the A, A'
and/or C groups are
substituted by halogen. In another embodiment, the A, A' and/or C groups are
substituted by F. In
another embodiment, the A, A' and/or C groups are substituted by Cl. In
another embodiment,
the A, A' and/or C rings are substituted by Br.
[00109] The substituents of these A, A' and/or C groups of formula I,
I(a), IV, IX, IX(a)
and XI are independently selected from the group of hydrogen (e.g., no
substitution at a
particular position), hydroxyl, an aliphatic straight- or branched-chain C1 to
C10 hydrocarbon,
alkoxy, haloalkoxy, aryloxy, nitro, cyano, alkyl-CN, halo (e.g., F, Cl. Br,
1), haloalkyl,
dihaloalkyl, trihaloalkyl, COOH, C(0)Ph, C(0)-alkyl, C(0)0-alkyl, C(0)H,
C(0)NH2, -
OC(0)CF3, OCH2Ph, amino, aminoalkyl, alkylamino, mesylamino, dialkylamino,
arylamino,
amido, NHC(0)-alkyl, urea, alkyl-urea, alkylamido (e.g., acetamide),
haloalkylamido, arylamido,
aryl, and C5 to C cycloalkyl, arylalkyl, and combinations thereof. Single
substituents can be
present at the ortho, meta, or para positions. When two or more substituents
are present, one of
them is preferably, though not necessarily, at the para position.
[00110] In one embodiment the B group of formula I, I(a), II, III, IV,
IVa and V is
selected from substituted or unsubstituted- thiazole, thiazolidine, oxazole,
oxazoline, oxazolidine,
benzene, pyrimidine, irnidazole, pyridine, furan, thiophene, isoxazole,
piperidine, pyrazole,
indole and isoquinoline, wherein said B ring is linked via any two positions
of the ring to X and
Y or directly to the A and/or C rings.
[00111] In one embodiment the B group of formula I, I(a), II, III, IV,
IVa and V is
unsubstituted. In another embodiment the B group of formula I, I(a), II, III,
IV, IVa and V is:
53
CA 3030689 2019-01-18
S-,;\ S
)--1µ.- 0
--µ 1,
- N ^ N /
'(thiazole), i-µN) (thiazole), H (thiazolidine), N /(oxazole),
0--A 41
---NN..r),. V(N),, = i
e (oxazoline), H (oxazolidine), -0-'
(benzene), (benzene),
HN'Y
)----=N -{-
-44 (pyrimidine), ''L=i- (imidazole), -r,'
(pyridine), 0 i's (furan),
_jc Q
---N -A. K-ts
ii (thiophene), "(isoxazole), .pr, , ;^' (piperidine),
,t'___J.:=Z-1
\-40CIµ i tc,..i....,..\:.N = .
5 rrt (pyrazole), ' N (indole), or ,--
'(isoquinoline).
= [00112] In another embodiment the B group of formula I, I(a), II,
III, IV, IVa and V is
substituted. In another embodiment the B group of formula I, I(a), II, III,
IV, IVa and V is:
Sf s...,..-'s...,..-'(Rio)i (R10)1
5 i s s-IA R
-4i-Ril ---- 7J-(FtioN t_i..õ , 11
N's-,s5 N-\ ' N"---/
NN/
(thiazole), R11 (thiazole), H (thiazolidine),
(R10)1
(R10)1
(:)/(Rio)1 01 A R11
P-1-1 R11
1--tt.7.F" l't.
e- (oxazole), N (oxazoline), H (oxazolidine),
(R101 (Rio)1 (R10)1
N-1- NW" si---1
E--(,, N
10 .,=-= (benzene), ¨ Ril (benzene) --'
(pyrimidine), -1.%, (imidazole),
(l10)1 (R10)1 0R10/1
5-Rii a
N-=-Kt--....t....711 (Rio), µ-g----/-
--ssi (furan), -Ns' (pyridine), il (thiophene), (isoxazole),
(R10)1 ,Pr'
IRIR11 h'(R10)1 '-7 1 -NH (Rio)! R1'
\-N R1 i.".."--=N
A=V-IN
(R10), R 'is µ01,VI-
(piperidine), 11 (pyrazole), ' N
(indole), or
JR"
1-CC1
..- N
-,1--= (isoquinoline);
54 -
CA 3030689 2019-01-18
=
wherein RI() and R11 are independently hydrogen, 0-alkyl, 0-haloalkyl, F, Cl,
Br, I, haloalkyl,
CF3, CN, -CH2CN, NH2, hydroxyl, -(CH2);NHCH3, -(CH2)1NH2, -(CH2)1N(CH3)2, -
0C(0)CF3,
C1-05 linear or branched alkyl, haloalkyl, alkylamino, aminoalkyl, -OCH2Ph, -
NHCO-alkyl,
COOH, -C(0)Ph, C(0)0-alkyl, C(0)H, -C(0)NH2 or NO2.
N ,s5
[00113] In another embodiment the B group is e
(thiazole). In another
=
embodiment the B group is N
(thiazole). In another embodiment the B group is
0
N
H (thiazolidine). In another embodiment the B group is N
(oxazole). hi another
0
embodiment the B group is ir
(oxazoline). In another embodiment the B group is
V(N
H (oxazolidine). In another embodiment the B group is --or(benzene).
In another
411 embodiment the B group is
(benzene). In another embodiment the B group is
= N
)=-N
====, (pyrimidine). In another embodiment the B group is
(imidazole). In another
embodiment the B group is
(pyridine). In another embodiment the B group is
(furan). In another embodiment the B group is
(thiophene). In another
O-N
_
embodiment the B group is sir (isoxazole). In another embodiment the B
group is
C-IN?
(piperidine). In another embodiment the B group is
(piperidine). In another
CA 3030689 2019-01-18
embodiment the B group is
/(pyrazole). In another embodiment the B group is
=, ,
1 tsts) (indole). In another embodiment the B group is
[001 14J In
one embodiment the B group of formula I, I(a), II, III, IV, TVa and V is
substituted by Rio and R11. In another embodiment, R10 and R11 are both
hydrogens. In another
embodiment, Rio and Rii are independently 0-alkyl. In another embodiment, Rio
and R11 are
independently 0-haloalkyl. In another embodiment, Rio and R11 are
independently F. In another
embodiment, Rio and R11 are independently Cl. In another embodiment, Rio and
Rii are
independently Br. In another embodiment, Rio and R11 are independently I. In
another
embodiment, Rio and R11 are independently haloalkyl. In another embodiment,
Rio and Rii are
independently CF3. In another embodiment, Rio and Rii are independently CN. In
another
embodiment, Rio and R11 are independently -CH2CN. In another embodiment, Rio
and R11 are
independently NH2. In another embodiment, Rio and R1, are independently
hydroxyl. In another
embodiment, Rio and Rii are Independently -(CH2)1NHCH3. In another embodiment,
Rio and Ri
are independently -(CH2)1NH2. In another embodiment, Rio and Ri I are
independently -
(CH2),N(CH3)2. In another embodiment, Rio and R11 are independently -0C(0)CF3.
In another
embodiment, Rio and R,1 are independently Ci-05 linear or branched alkyl. In
another
embodiment, Rio and R 1 i are independently C1-05 linear or branched
haloalkyl. In another
embodiment, Rio and R11 are independently C1-05 linear or branched alkylamino.
In another
embodiment, Rio and R11 are independently C1-05 linear or branched aminoalkyl.
In another
embodiment, Rio and RI are independently -OCH2Ph. In another embodiment, Rio
and Rii are
independently -NHCO-alkyl. In another embodiment, Rio and R1, are
independently COOH. In
another embodiment, Rio and Rii are independently -C(0)Ph. In another
embodiment, Rio and
are independently C(0)0-alkyl. In another embodiment, Rio and R,1 are
independently
C(0)H. In another embodiment, Rio and R11 are independently -C(0)N112. In
another
embodiment, Rio and 1211 are independently NO2.
[00115] In
another embodiment the B group of formula I, I(a), II, III, IV, IVa and V is
(R-10)1
=
5 Ska
Rit
(thiazole), wherein Rio and R11 are independently H and 1 is I. In another
56
CA 3030689 2019-01-18
embodiment, R10 and RI I are independently 0-alkyl. In another embodiment, Rio
and R1 are
independently 0-haloalkyl. In another embodiment, Rio and R11 are
independently F. In another
embodiment, R10 and Rii are independently Cl. In another embodiment, Ri0 and
Ri are
independently Br. In another embodiment, R10 and Rii are independently L In
another
embodiment, R10 and R11 are independently haloalkyl. In another embodiment,
R10 and R11 are
independently CF3. In another embodiment, Rio and 1211 are independently CN.
In another
embodiment, R10 and R11 are independently -CH2CN. In another embodiment, Rio
and R11 are
independently NH2. In another embodiment, Rio and Rii are independently
hydroxyl. In another
embodiment, Rio and R11 are independently -(CH2)1NHCH3. In another embodiment,
R10 and R11
are independently -(CH2)1llf12. In another embodiment, RIO and RH are
independently - '
(CH2)1N(CH3)2. In another embodiment, Rio and RH are independently -0C(0)CF3.
In another
embodiment, R10 and R11 are independently C1-05 linear or branched alkyl. In
another
embodiment, R10 and R11 are independently C1-05 linear or branched haloalkyl.
In another
embodiment, R10 and 1211 are independently C1-05 linear or branched
alkylamino. In another
embodiment, RI0 and 1211 are independently CI-05 linear or branched
aminoalkyl. In another
embodiment, R10 and R11 are independently -OCH2Ph. In another embodiment, Ri0
and R11 are
independently -NHCO-alkyl. In another embodiment, Rio and R11 are
independently COOH. In
another embodiment, Rio and R11 are independently -C(0)Ph. In another
embodiment, R10 and
R11 are independently C(0)0-alkyl. In another embodiment, R10 and Rii are
independently
C(0)H. In another embodiment, Rio and R11 are independently -C(0)NH2. In
another
embodiment, R10 and R11 are independently NO2.
[00116] In another embodiment the B group of formula I, I(a), II, III,
IV, IVa and V is
Ril
)=--KI1R10)1
(imidazole), wherein Rio and R11 are independently H and 1 is 1. In another
embodiment, R10 and R11 are. independently 0-alkyl. In another embodiment, R10
and R11 are
independently 0-haloalkyl. In another embodiment, Rio and R11 are
independently F. In another
embodiment, R10 and R11 are independently Cl. In another embodiment, R10 and
RI, are
independently Br. In another embodiment, Rio and R,1 are independently I. In
another
embodiment, R10 and R11 are independently haloalkyl. In another embodiment,
R10 and R11 are
independently CF3. In another embodiment, Rio and R11 are independently CN. In
another
57
CA 3030689 2019-01-18
embodiment, Rio and R11 are independently. -CH2CN. In another embodiment, R10
and R11 are
independently NH2. In another embodiment, R10 and R11 are independently
hydroxyl. In another
embodiment, Rio and Rii are independently 4CH2)1NHCH3. In another embodiment,
Rio and
are independently -(CH2)1NH2. In another embodiment, R10 and R11 are
independently -
(CH2)1N(CH3)2, In another embodiment, Rio and R11 are independently -0C(0)CF3.
In another
embodiment, R10 and R11 are independently C1-05 linear or branched alkyl. In
another
embodiment, Rio and R11 are independently CI-Cs linear or branched haloalkyl.
In another
embodiment, Rio and Ri are independently CI-Cs linear or branched alkylamino.
In another
embodiment, Rio and Rii are independently C1-05 linear or branched aminoalkyl.
In another
embodiment, R10 and R11 are independently -OCH2Ph. In another embodiment, R10
and R11 are
independently -NHCO-alkyl. In another embodiment, Rio and R11 are
independently COOH. In
another embodiment, Rio and R11 are independently -C(0)Ph. In another
embodiment, Rio and
R11 are independently C(0)0-alkyl. In another embodiment, Rio and R11 are
independently
C(0)1-I. In another embodiment, RID and R11 are independently -C(0)NH2. In
another
embodiment, Rio and R11 are independently NO2-
[001 17] In another embodiment the B group of formula I, I(a), II, III,
IV, IVa and V is
(1110)1\,... R,,
(isoquinoline), wherein Rio and R11 are independently H and 1 is 1. In another
embodiment, Rio and R11 are independently 0-alkyl. In another embodiment, Rio
and R11 are '
independently 0-haloalkyl. In another embodiment, Rio and Ri are independently
F. In another
embodiment, R10 and R11 are independently Cl. In another embodiment, R10 and
R11 are
independently Br. In another embodiment, R10 and RI are independently I. In
another
embodiment, Rio and R11 are independently haloalkyl. In another embodiment,
Rio and R11 are
independently CF3. In another embodiment, Rio and R11 are independently CN. In
another
embodiment, Rio and R11 are independently -CH2CN. In another embodiment, Rio
and 1211 are
independently NI-12. In another embodiment, Rio and 1211 are independently
hydroxyl. In another
embodiment, R10 and R11 are independently -(CH2)1NHCH3. In another embodiment,
Rio and Rii
are independently -(CH2),NH2. In another embodiment, Rio and R11 are
independently -
(CH2),N(CH3)2. In another embodiment, R10 and R11 are independently -0C(0)CF3.
In another
embodiment, Rio and R11 are independently C1-05 linear or branched alkyl. In
another
embodiment, R10 and R,1 are independently C1-05 linear or branched haloalkyl.
In another
58
CA 3030689 2019-01-18
embodiment, Rio and R11 are independently C1-05 linear or branched alkylamino.
In another
embodiment, Rio and R,1 are independently C,-05 linear or branched aminoalkyl.
In another
embodiment, Rio and R11 are independently -OCH2Ph. In another embodiment, Rio
and R11 are
independently -NHCO-alkyl. In another embodiment, Rio and 11.11 are
independently COOH. In
another embodiment, Rio and Rii are independently -C(0)Ph. In another
embodiment, R10 and
R are independently C(0)0-alkyl. In another embodiment, R10 and R1, are
independently
C(0)H. In another embodiment, Rio and R11 are independently -C(0)NH2. In
another
embodiment, Rio and R11 are independently NO2.
[00118] In one embodiment, the X bridge of formula I, Ia, H, LH, IV, IVa and
XI is a bond. In
another embodiment, the X bridge is NH. In another embodiment, the X bridge is
Ci to C5
hydrocarbon. In another embodiment, the X bridge is CH2. In another
embodiment, the X bridge is ¨
CH2-CH2-. In another embodiment, the X bridge is 0. In another embodiment, the
X bridge is S.
[00119] In one embodiment, the Y bridge of formula I, Ia, II, HI, IV, IVa, VI,
and VII is C.O.
In another embodiment, the Y bridge is C=-S. In another embodiment, the Y
bridge is C=N(NH2)-=
In another embodiment, the Y bridge is -C=NOH. In another embodiment, the Y
bridge is ¨CH-OH.
In another embodiment, the Y bridge is -C---CH-(CN). In another embodiment,
the Y bridge is -
C=N(CN). In another embodiment, the Y bridge is -C=C(CH3)2. In another
embodiment, the
bridge is -C=N-0Me. In another embodiment, the Y bridge is -(C_0)NH-. In
another embodiment,
the Y bridge is -NH(C=0)-. In another embodiment, the Y bridge is ¨(C=0)-0. In
another
embodiment, the Y bridge is ¨0-(C=0). In another embodiment, the Y bridge is
¨(CH2)1_5-(C=0).
In another embodiment, the Y bridge is ¨(C=0)-(0-12)1-5- In another
embodiment, the Y bridge is-S.
In another embodiment, the V bridge is SO. In another embodiment, the Y bridge
is SO2. In another
embodiment, the Y bridge is In another embodiment, the Y bridge is ¨(S02)-
NH-. In
- another embodiment, the Y bridge is ¨NH-(S02)-=
[00120] In one embodiment, Ri, R2, R3, R4, Rs and R6 of formula In, II, HI,
IV, IV(a), V, VI,
VIII, IX, IX(a), XI(a), Xl(b), Xl(c), XI(d) and XI(e) are independently
hydrogen. In another
embodiment, RI, R2, RS, R4, RS and R6 are independently 0-alkyl. In another
embodiment, RI, R2,
R3, 114, Rs and R6 are independently 0-haloalkyl. In another embodiment,.121,
R2, R3, R4, Rs and R6
are independently F. In another embodiment, Ri, R2, R3, R4, Rs and R6 are
independently Cl. In
59
=
CA 3030689 2019-01-18
another embodiment, Rit, R2, R3, Ra, Rs and R6 are independently Br. In
another embodiment, R1,
R2, R3, 114, R5 and R6 are independently I. In another embodiment, Ri, R2, R3,
R4, R5 and R6 are
independently haloalkyl. In another embodiment, RI, R2, R3, R4, R5 and R6 are
independently CF3.
In another embodiment, 121, Ri, R3, itt, Rs and R6 are independently CN. In
another embodiment,
RI, R2, R3, R4, R5 and R6 are independently -CH2CN. In another embodiment, RI,
R2, R3, R4, R5
and R6 are independently NI-12. In another embodiment, Ri, R2, R3, R4, Rs and
R6 are independently
hydroxyl. In another embodiment, RI, R2, R3, It, R5 and R6 are independently -
(CH2)NHC1-13. In
another embodiment, R3, R2, R3, R4, R5 and R6 are independently -(CH2)1NH2. In
another
embodiment, R1, R2, R3, R.4, R5 and R6 are independently -(CH2)1N(CH3)2. In
another embodiment,
Ri, R2, R3, R4, R5 and R6 are independently -0C(0)CF3. In another embodiment,
RI, R2, R3, 124, Rs
and R6 are independently C1-05 linear or branched alkyl. In another
embodiment, RI, R2, R3, R4, R5
and R6 are independently haloalkyl. In another embodiment, RI, R2, R3, R4, Rs
and R6 are
independently alkylamino. In another embodiment, RI, R2, R3, R4, R5 and R6 are
independently
aminoalkyl. In another embodiment, RI, R2, R3, R4, Rs and R6 are independently
-OCH2Ph. In
another embodiment, Ri, R2, RI R4p R5 and R6 are independently -I\THCO-alkyl.
In another
embodiment, RI, R2, R3, R4, R5 and Li are independently COOH. In another
embodiment, Ri, R2,
R3, Ra, R5 and R6 are independently -C(0)Ph. In another embodiment, R1, R2,
R3, 124, R5 and R6 are
independently C(0)0-alkyl. In another embodiment, RI, R2, R3, 114, R5 and R6
are independently
C(0)H. In another embodiment, RI, R2, R3, R4, R5 and R6 are independently -
C(0)NH2. In another
embodiment, RI, R2, R3, R4, R5 and R6 are independently NO2.
[00121) In one embodiment, this invention is directed to a compound of formula
XII:
(R4)n
/ p
R5IN
(Xl)
.. wherein,
P and Q are independently H or
* (R)),.
R2
=
CA 3030689 2019-01-18
W is C=0, C=S, SO2 or S=0;
wherein at least one of Q or P is not hydrogen;
R1 and R4 are independently H, 0-alkyl, I, Br, Cl, F, alkyl, haloalkyl,
aminoalkyl, OCH2Ph, OH,
CN, NO2, -NHCO-alkyl, COOH, -(CH2)1NHCH3, -(CH2),NH2, -(C112);N(CH3)2; C(0)0-
alkyl or
C(0)H; wherein at least one of R1 and R4 is not hydrogen;
R2 and R5 are independently H, 0-alkyl, I, Br, Cl, F, alkyl, haloalkyl,
aminoalkyl, OCH2Ph, OH,
CN, NO2, -NHCO-alkyl, COOH, C(0)0-alkyl or C(0)H;
m is an integer between 1-4;
i is an integer between 0-5; and
=
n is an integer between 1-4;
or its pharmaceutically acceptable salt, hydrate, polymorph, metabolite,
tautomer or isomer.
[00122] In one embodiment, this invention is directed to a compound of formula
XIII:
(R4):
I
R5 NH
R2
wherein
Z is 0 or S;
R1 and R4 are independently H, 0-alkyl, I, Br, Cl, F, alkyl, haloalkyl,
aminoalkyl, OCH2Ph, OH,
CN, NO2, -NHCO-alkyl, haloalkyl, aminoalkyl, -(CH2)1NHCH3, -(C112)1NH2, -
(CH2),N(CH3)2 ;
COOH, C(0)0-alkyl or C(0)H; wherein at least one of RI and R4 is not hydrogen;
Rz and R5 are independently H, 0-alkyl, I. Br, Cl, F, alkyl, haloalkyl,
aminoalkyl, -(CH2)1NHCH3, -
(CH2)1NI-12, -(CH2);N(CH3)2; OCH2Ph, OH, CN, NO2, -NHCO-alkyl, COOH, C(0)0-
alkyl or
C(0)H;
m is an integer between 1-4;
i is an integer between 0-5; and
n is an integer between 1-4;
or its pharmaceutically acceptable salt, hydrate, polymorph, metabolite,
tautomer or isomer,
61
CA 3030689 2019-01-18
[00123] In one embodiment, this invention is directed to a compound of formula
XIV:
0
014)n
R5 NH
R2
(XIV)
wherein It1 and R4 are independently H, 0-alkyl, I, Br, Cl, F, alkyl,
haloalkyl, aminoalkyl, -
(CH2)1NHCH3, -(CH2)1NII2, -(CH2)N(CH3)2, OCH2Ph, OH, CN, NO2, COOH,
C(0)0-alkyl or C(0)H; wherein at least one of Ri and R.4 is not hydrogen;
R2 and R5 are independently 'H, 0-alkyl, I, Br, Cl, F, alkyl, haloalkyl,
aminoalkyl, OCH2Ph, OH,
CN, NO2, -NHCO-alkyl, COOH, C(0)0-alkyl or C(0)H;
m is an integer between 1-4;
i is an integer between 0-5; and
n is an integer between 1-4;
or its pharmaceutically acceptable salt, hydrate, polymorph, metabolite,
tautomer or isomer.
[00124] In one embodiment, R1 of compound of formula XII, XIII and XIV is
OCH3. In another
embodiment, R1 of compound of formula XII, mu and XIV is 4-F. In another
embodiment, R1 of
compound of formula XII, XIII and XIV is OCH3 and m is 3. In another
embodiment, R4 of
compound of formula XII, XIII and XIV is 4-F. In another embodiment, R4 of
compound of
formula XII, XIII and XIV is OCH3. In another embodiment, R4 of compound of
formula XIV is
CH3. In another embodiment, R4 of compound of formula XII, XIII and XIV is 4-
Cl. In another
embodiment, R4 of compound of formula XII, XIII and XIV is 4-N(Me)2. In
another embodiment,
R4 of compound of formula XII, XIII and XIV is OBn. In another embodiment, R4
of compound of
formula XII, XM and XIV is 4-Br. In another embodiment, R4 of compound of
formula XII, XIII
and XIV is 4-CF3. Non limiting examples of compounds of formula XIV are
selected from: (2-
phenyl-If/4mi dazol -4-yl)(3,4,5-trimethox yphenyl)methanone (12aa), (4-
fluorophenyl)(2-Phenyl-
1l1-imidazol-4-ypmethanone (12af), (2-(4-
fluoropheny1)-1H-imidazol-4-y1)(3,4,5-
trimethoxyphenyl)methanone (12ba), (2-(4-
methoxypheny1)-1H-imidazol-4-y1)(3,4,5-
trimethoxyphenyOmethanone (12ca), (4-fluorophenyl)(2-(4-methoxyphenyl)-1/1-
imidazol-4-
yl)methanone (12cb), (2-(p-toly1)-1H-imidazol-4-y1)(3,4,5-
trimethoxyphenypmethanone (12da), (4-
62
CA 3030689 2019-01-18
fluoropherry1)(2-(p-toly1)-1H-imidazol-4-y1)methanone (12db), (4-
hydrox y-3,5-
dimethox ypheny1)(2-(p-toly1)-1H-imidazol-4-yOmethanone
(12dc), (2-(4-chloropheny1)-IH-
imidazol-4-y1)(3,4,5-trimetboxyphenyl)methanone (121a), (2-(4-chloropheny1)-1H-
imidazol-4-y1)(4-
fluorophenyl)methanone (12113), (2-(4-
chloropheny1)-1H-imidazol-4-y1)(4-hydrox y-3,5-
dimethoxyphenyl)methanone (12fc), (2-(4-(dimethylamino)pheny1)-1H-imidazol--4-
y1)(3,4,5-
trimethox yphenyl)metharione (12ga); (2-(4-
(dimethylamino)pheny1)-1H-imidazol-4-y1)(4-
fluorophenyl)methanone (12gb), (2-
(3,4-dimethoxypheny1)-1H-imidazol-4-y1)(3,4,5-
trimethoxyphenyOmethanone (12ha), (2-(4-
(benzyloxy)pheny1)-1H-imidazol-4-y1)(4-
fluorophenyl)methanone (12jb), (2-(4-
bromopheny1)-1H-imidazol-4-y1)(3,4,5-
trimethox yphenyl)methanone (121a),
(2-(4-(trifluoromethyl)pheny1)-1H-imidazol-4-y1)(3,4,5-
trimethox yphenypmethanone (12pa).
[00125] In one embodiment, this invention is directed to a compound of formula
XIVa:
0
(R4)n
/ (RILI
R5
R2
R9
(XlVa)
wherein R1 and Rs are independently H, 0-alkyl, I, Br, Cl, F, alkyl,
haloalkyl, aminoalkyl, -
(CH2)1NHCH3, -(C1-12)1NH2, -(CH2)1N(CH3)2, OCH2Ph, OH, CN, NO2, -NHCO-alkyl,
COOH,
C(0)0-alkyl or C(0)H; wherein at least one of R1 and R4 is not hydrogen;
R2 and R5 are independently H, 0-alkyl, I, Br, Cl, F, alkyl, haloalkyl,
aminoalkyl, -(CH2)1NHCH3, -
(CH2)iNH2, -(CH2),N(CH3)2, OCH2Ph, OH, CN, NO2, -NECO-alkyl, COOH, C(0)0-alkyl
or
C(0)H;
129 is H, linear or branched, substituted or unsubstituted alkyl, substituted
or unsubstituted aryl,
CH2Ph, substituted benzyl, haloalkyl, aminoalkyl, OCH2Ph, substituted or
unsubstituted S02-Aryl,
substituted or unsubstituted -(C=0)-Aryl or OH;
wherein substitutions are independently selected from the group of -hydrogen
(e.g., no substitution at
a particular position), hydroxyl, an aliphatic straight- or branched-chain C1
to C10 hydrocarbon,
alkoxy, haloalkoxy, aryloxy, nitro, cyano, alkyl-CN, halo (e.g., F, Cl, Br,
1), haloalkyl, dihaloalkyl,
trihaloalkyl, COOH, C(0)Ph, C(0)-alkyl, C(0)0-alkyl, C(0)H, C(0)N1-12, -
0C(0)CF3, OCH2Ph,
amino, aminoalkyl, alkylarnino, mesylamino, dialkylamino, arylamino, amido,
NHC(0)-alkyl, urea,
63
CA 3030689 2019-01-18
alkyl-urea, alkylarnido (e.g., acetarnide), haloallcylamido, arylamido, aryl,
and C5 to C7 cycloalkyl,
arylalkyl, and combinations thereof;
m is an integer between 1-4;
i is an integer between 0-5 ; and
n is an integer between 1-4;
or its pharmaceutically acceptable salt, hydrate, polymorph, metabolite,
tautomer or isomer.
[00126] In one embodiment, R9 of compound of formula XIVa is CH3. In another
embodiment, =
R9 of compound of formula XIVa is CH2Ph_ In another embodiment, R9 of compound
of formula
XIVa is (S02)Ph. In another embodiment, R9 of compound of formula XIVa is
(S02)-Ph-OCH3. In
another embodiment, R9 of compound of formula XIVa is H. In another
embodiment, 124 of
compound of formula XIVa is H. In another embodiment, R4 of compound of
formula XIVa is
CH3. In another embodiment, R4 of compound of formula XIVa is OCH3. In another
embodiment,
R4 of compound of formula XlVa is OH. In another embodiment, R4 of compound of
formula
XIVa is 4-Cl. In another embodiment, R4 of compound of formula XIVa is 4-
N(Me)2. In another
embodiment, R4 of compound of formula XIVa is OBn. In another embodiment, it1
of compound
of formula XIVa is OCH3; m is 3 and R2 is H. In another embodiment, R1 of
compound of formula
XIVa is F; m is I and R2 is H. Non limiting examples of compounds of formula
XIVa are selected
from: (4-fluorophenyl)(2-phenyl-1-(phenylsulfony1)-1H-imidazol-4-yl)methanone
(liar), (4-
fluorophenyl)(2-(4-methoxypheny1)-1-(phenylsulfonyl)-1H-imidazol-4-
y1)methanone (11cb), (4-
fluorophenyl)(1-(phenyl sulfony1)-2-(p-toly1)- I H-imidazol-4-ypmethanone
(11db), (2-(4-
chloropheny1)-1-(phenylsulfony1)-1H-imidaz,o1-4-y1)(4-fluorophenyl)methanone
(11fb), (244-
(dimethylami no)pheny1)-1 -(pheny1sulfony1)-1H-imidazol-4-y1)(3,4,5-
trimethoxyphenypmethanone
(11ga), (2-(4-
(dimethylamino)pheny1)- I -(phenylsulfony1)-1H-imidazol-4-y1)(4-
fluorophenyl)methanone (11gb), (2-(3,4-dimethox ypheny1)-1-(phenylsulfony1)-1
H-im idazol-4-
yl)(3,4,5-trimethoxyphenyl)methanone (11ha), (2-(4-(benzyloxy)pheny1)-1-
(phenylsulfony1)-1H-
imidazol-4-y1)(4-fluorophenyl)methanone (11jb),
(2-(4-(dimethylarnino)pheny1)-14(4-
methoxyphenyl)su 'fon y1)-1H-imidazol-4-y1)(4-fl uorophenyl)methanone (12gba),
(1 -benzy1-2-(p-
tol yl)-1H-i m idazol-4-y1)(3,4,5-trimethox yphen yl)m eth anon e (12daa), (1-
meth y1-2-(p-toly1)-1 H-
imidazol-4-y1)(3,4,5-trimethoxyphenyl)methanone
(12dab), .. (4-fluorophenyl)(2-(4-
methoxypheny1)-1-methyl-1H-irnidazol-4-yl)methanone (12cba).
[00127] In one embodiment, this invention is directed to a compound of formula
XV:
64
CA 3030689 2019-01-18
0
(Itt)n OCH3
/
R5 NH OCH3
OCH3
(XV)
wherein R4 and R5 are independently H, 0-alkyl, I, Br, Cl, F, alkyl,
haloalkyl, aminoalkyl, -
(CH2),NHCH3, -(CH2)1NH2, -(CH2)1N(CH3)2, OCH2Ph, OH, CN, NO2, -NHCO-alkyl,
COOH,
C(0)0-alkyl or C(0)H;
i is an integer between 0-5; and
n is an integer between is 1-4;
or its pharmaceutically acceptable salt, hydrate, polymorph, metabolite,
tautomer or isomer.
[00128] In one embodiment, R4 of compound of formula XV is H. In another
embodiment, R4 of
compound of formula XV is F. In another embodiment, R4 of compound of formula
XV is Cl. In
another embodiment, I14 of compound of formula XV is Br. In another
embodiment, R4 of
compound of formula XV is I. In another embodiment, R4 of compound of formula
XV is N(Me)2-
In another embodiment, R4 of compound .of formula XV is OBn. In another
embodiment, R4 of
compound of formula XV is OCH3. In another embodiment, R4 of compound of
formula XV is
CH3. In another embodiment, R4 of compound of formula XV is CF3. Non limiting
examples Of
compounds of formula XV are selected from: (2-
phenyl- 1H-imidazol-4-y1)(3,4,5-
trimethoxyphenypmethanone (12aa), (2-(4-
fluorophenyI)- I H-imidazol-4-y1)(3,4,5-
trimethoxyphenyl)methanone (12ba), (2-(4-
methox ypheny1)- I H-im idazol-4-y1)(3,4,5-
trimethox yphenyl)methanone (12ca), (2-(p-tolyI)- I H-imidazol-4-
y1)(3,4,5-
trimethoxyphenyl)methanone (12da), (3,4,5-trimethoxyphenyl)(2-(3,4,5-
trimethoxypheny1)-1H-
imidazol-4-yDrnethanone (12ea), (2-(4-
chloropheny1)-1H-imidazol-4-y1)(3,4,5-
n-i methox yp hen yl )met hanone
(12fa), (2-(4-(dimethylamino)pheny1)-1H-imidazol-4-y1)(3,4,5-
trimethox yphenyl)methanone (12ga), (2-(3
,4-dimethoxyphen y1)-4 H-imidazo1-4-y1)(3,4,5-
trimethox yphenyl)methanone (12ha), (2-(2-(trifl uoromethyl)phen y1)- 1H-
imidazol-4-y1)(3,4,5-
tri methox yphenypmethanone (12ia), (2-(4-
(benzylox y)phen y1)-1H-imidazol-4-y1)(3,4,5-
trimethoxyphenypmethanone (12ja), (2-(4-
hydroxypheny1)-1H-imidazol-4-y1)(3,4,5-
trimethoxyphenyOmethanone (12ka), (2-(4-
bromophenyI)-1H-imidazol-4-y1)(3,4,5-
CA 3030689 2019-01-18
trimethox yphen yl)methanone
(121a), (2-(4-(trifluoromethyl)pheny1)-1H-imidazol-4-y1)(3,4,5-
trimethoxyphenyl)methanone (12pa).
[00129] In one embodiment, this invention is directed to a compound of formula
XVI:
0
(R4)n =
/ I
R5 R3
(XVI)
wherein R4 and Rs are independently H, 0-alkyl, I, Br, Cl, F, alkyl,
haloalkyl, aminoalkyl, -
(CH2)1NHCH3, -(CH2)1NH2, -(CH2)1N(CH3)2, OCH2Ph, OH, CN, NO2, -NHCO-alkyl,
COOH,
C(0)0-alkyl or C(0)H;
R3 is 1, Br, Cl, or F;
i is an integer between 0-5; and
n is an integer between 1-4;
or its pharmaceutically acceptable salt, hydrate, polymorph, metabolite,
tautomer or isomer.
[00130] In one embodiment, R3 of compound of formula XVI is halogen. In
another
embodiment, R3 is F. In another embodiment, R3 is a In another embodiment R3
is Br. In another
embodiment R3 is I. In another embodiment R4 is H. In another embodiment R4 is
OCH3. In
another embodiment R4 is 0CH3; n is 3 and R5 is H. In another embodiment R4 is
CH3. In another
embodiment R4 is F. In another embodiment R4 is Cl. In another embodiment R4
is Br. In another
embodiment 114 is I. In another embodiment R4 is N(Me)2. In another embodiment
R4 is OBn. In
another embodiment, R3 is F; R5 is hydrogen; n is 1 and R4 is 4-Cl. In another
embodiment, R3 is F;
R5 is hydrogen; n is I and R4 is 4-0CH3. In another embodiment, R3 is F; R5 is
hydrogen; n is 1 and
R4 is 4-CH3. In another embodiment, R3 is F; R5 is hydrogen; n is 1 and R4 is
4-N(Me)2. In another
embodiment, R3 is F; Rs is hydrogen; n is 1 and R4 is 4-0Bn. Non limiting
examples of compounds
of formula XVI are selected from: (4-fluorophenyl)(2-phenyl-1H-imidazol-4-
yl)methanone (12a1),
(4-fl uorophenyl)(2-(4-methox ypheny1)-1H-imidazol-4-yOmethanone (12cb), (4-
fluorophenyl)(2-(p-
toly1)-1H-imidazol-4-yl)methanone (12db), 4-fluorophenyl)(2-(3,4,5-
trimethoxypheny1)- 1 H-
imidazol-4-yl)methanone (12eb), (2-(4-
chloropheny1)-1H-imidazol-4-y1)(4-
fl uorophenypmethanone (1211,), (2-(4-
(di methylami no)phen yl)-1H-i midazol-4-y1)(4-
66
CA 3030689 2019-01-18
fluorophenypmethanone (12gb), (2-(4-
(benzyloxy)pheny1)-1H-imidazol-4-y1)(4-
fluorophenypmethanone (12jb).
[00131] In one embodiment, this invention is directed to a compound of formula
XVII:
0
NH
R2
(XVII)
wherein 144 is H, 0-alkyl, 1, Br, Cl, F, . alkyl, haloalkyl, aminoalkyl,
OCH2Ph, OH, CN, NO2, -
NHCO-alkyl, COOH, C(0)0-alkyl or C(0)H;
wherein R1 and R2 are independently H, 0-alkyl, I, Br, Cl, F, alkyl,
haloalkyl, aminoaLkyl,
OCH2Ph, OH, CN, NO2, -NHCO-alkyl, COOH, C(0)0-alkyl or C(0)H;
and
m is an integer between 1-4;
or its pharmaceutically acceptable salt, hydrate, polymorph, metabolite,
tautomer or isomer.
In one embodiment, 114 of compound of formula XVII is halogen. In another
embodiment, 114 is F.
In another embodiment, 114 is Cl. In another embodiment 144 is Br. In another
embodiment 114 is I.
In another embodiment, 144 is OCH3. In another embodiment, 114 is CH3. In
another embodiment,
R. is N(Me)2. In another embodiment, 124 is CF3. In another embodiment, R4 is
OH. In another
embodiment, 114 is OBn. In another embodiment, R1 of compound of formula XVII
is halogen. In
another embodiment, R1 of compound of formula XVII is F. In another
embodiment, R1 of
compound of formula XVII is Cl. In another embodiment, R1 of compound of
formula XVII is Br.
In another embodiment, R1 of compound of formula XVII is I. In another
embodiment, R1 of
compound of formula XVII is OCH3. In another embodiment, R1 of compound of
formula XVII is
OCH3, m is 3 and R2 is H. In another embodiment, R1 of compound of formula
XVII is F, m is 1
and R2 is H. In another embodiment, R4 is F; R2 is hydrogen; n is 3 and R1 is
()CHI In another
embodiment, R4 is OCH3; R2 is hydrogen; n is 3 and RI is OCH3. In another
embodiment, 114 is
CH3; R2 is hydrogen; n is 3 and R1 is OCH3. In another embodiment, 144 is Cl;
R2 is hydrogen; n is 3
and R1 is OCH3 In another embodiment, 144 is N(Me)2; R2 is hydrogen; n is 3
and R1 is OCH3. In
one embodiment, R4 of compound of formula XVII is halogen, R1 is H and R2 is
halogen. In one
embodiment, Ri of compound of formula XVII is halogen, R1 is halogen and R2 is
H. In one
67
CA 3030689 2019-01-18
embodiment, 114 of compound of formula XVII is alkoxy, R1 is halogen and R2 is
H. In one
embodiment, 114 of compound of formula XVII is methoxy, R1 is halogen and R2
is H. Non
limiting examples of compounds of formula XVII are selected from: (2-(4-
fluoropheny1)-1H-
imidazol-4-y1)(3,4,5-trimethoxyphenyl)methanone (12ba), (2-(4-methox ypheny1)-
1H-imidazol-4-
yl)(3,4,5-trimethoxyphenyl)methanone (12ea), (4-fluorophenyl)(2-(4-
methox yphen y1)- 1 H-
imidazol-4-yl)methanone (.12eb), (2-(p-
to ly1)-1H-imidazol-4-y1)(3,4,5-
trimethox yp hen yOmeth anone (12da), (4-fluorophenyl)(2-(p-toly1)-1H-
inaidazol-4-yl)methanone
(12db), (4-Hydroxy-3,5-dimethoxyphenyl)(2-(p-toly1)-1H-imidazol-4-yOmethanone
(12de), (2-(4-
chloropheny1)-1H-irnidazol-4-y1)(3,4,5-trimethoz yphenypmethanone (12fa), (2-
(4-chlorophenyI)-
1H-imidazol-4-y1)(4-fluorophenyl)methanone (1M), (2-(4-chloropheny1)-1H-
imidazol-4-y1)(3,4,5-
trihydrox yphenyl)methanone (13fa), (2-(4-
(dimethylamino)pheny1)-1H-imidazol-4-y1)(3,4,5-
trimethoxyphenypmethanone (12ga), (2-(4-
(dimethylamino)pheny1)-1H-i midazol-4-y1)(4-
= fluorophenyl)methanone (12gb),
(2-(4-(benzyloxy)pheny1)-1H-imidazol-4-y1)(4-
fluorophenyl)methanone (12jb), (2-(4-h
ydrox yphen y1)-1H- imidazol-4-y1)(3,4,5-
trimethoxyphen yl)methanone (12ka), (2-(4-bromopheny1)-1H-
imidazol-4-y1)(3,4,5-
trimethoxyplienyl)methanone (121a), (2-(4-
(trifluoromethyl)pheny1)-1H-imidazol-4-y1)(3,4,5-
trimethoxyphen yOmethanone (12pa).
[00132] In another embodiment a compound of formula XVII is represented by the
structure of
formula 121b:
0
HN
CI (12M).
[00133] In another embodiment a compound of formula XVII is represented by the
structure of
formula 12cb:
0
HN
=
F
H3C0 (126).
[00134] In one embodiment, this invention is directed to a compound of formula
XVIII:
68
CA 3030689 2019-01-18
(R4L
/
R5
R8
(R7)q
(XVIII)
wherein
W is C=0, C=S, SO2 or S.---0;
R4 and R7 are independently H, 0-alkyl, I, Br, CI, F, alkyl, haloalkyl,
aminoalkyl, -(CH2)1NHCH3, -
(CH2)1NH2, -(CI-12)1N(CH3)2, OCH2Ph, OH, CN, NO2, -NICO-alkyl, COOH, C(0)0-
alkyl or
C(0)H;
R5 and R8 are independently H, 0-alkyl, I, Br, Cl, F, alkyl, haloalkyl,
aminoalkyl, -(CH2)1NI-ICH3, -
(CH2)1NH2, -(CI-12)1N(CH3)2, OCH2Ph, OH, CN, NO2, -NHCO-alkyl, COOH, C(0)0-
alkyl or
C(0)H;
n is an integer between 1-4;
i is an integer between 0-5; and
q is an integer between 1-4;
or its pharmaceutically acceptable salt, hydrate, polymorph, metabolite,
tautomer or isomer.
[00135] In one embodiment, W of compound of formula XVIII is C=0. In another
embodiment,
W of compound of formula XVIII is S02. In another embodiment, R4 of compound
of formula
XVIII is H. In another embodiment. R4 of compound of formula XVIII is NO2. In
another
embodiment, R4 of compound of formula XVIII is OBn. In another embodiment, R7
of compound
= 20 of formula XVIII is H. In another embodiment, R7 of compound of
formula XVIII is OCH3. In
another embodiment, R7 of compound of formula XVIII is OCH3 and q is 3. Non
limiting
examples of compounds of formula XVII are selected from: (4-methoxyphenyl)(2-
pheny1-1H-
imidazol-1-yl)methanone (12aba), (2-
phenyl-1H-imidazol- 1-y1)(3,4,5-
tri methox yphenyl)methanone (12aaa), 2-phenyl-1-(phenylsulfony1)-1H-
irnidazole (10a), 2-(4-
nitropheny1)-1-(phenylsulfony1)-1H-imidazole (10x), 2-(4-(benzyloxy)pheny1)-1-
(phenylsulfonyl)-
I H-imidazole (10j).
69
CA 3030689 2019-01-18
[00136] In one embodiment, this invention is directed to a compound of formula
XIX:
0 =
(R4)n
R5
1 R2
R8
(R7)q
(XIX)
wherein
W is C=0, C=S, SO2, S=0;
R1, R4 and R7 are independently H, 0-alkyl, I, Br, Cl, F, alkyl, haloalkyl,
aminoalkyl, -
(CH2);NHCH3, -(CH2)1NH2, -(CH2);N(CH3)2, OCH2Ph, OH, CN, NO2, -NHCO-alkyl,
COOH,
C(0)0-alkyl or C(0)H;
R2, R5 and Rs are independently H, 0-alkyl, 1, Br, Cl, F, alkyl, haloalkyl,
aminoalkyl, -
(CH2)1NHCH3, -(CH2)1NH2, -(CH2)1N(CH3)2, OCH2Ph, OH, CN, NO2, -NHCO-alkyl,
COOH,
C(0)0-alkyl or C(0)H;
m is an integer between 1-4;
n is an integer between 1-4;
i is an integer between 0-5; and
q is 1-4;
or its pharmaceutically acceptable salt, hydrate, polymorph, metabolite,
tautomer or isomer.
[00137] In one embodiment, RI, R4 and R7 of formula XIX are independently H.
In another
embodiment, RI, R4 and R7 of formula XIX are independently 0-alkyl. In another
embodiment, RI,
R4 and R7 of formula XIX are independently halogen. In another embodiment, R1,
R4 and 1117 of
formula XIX are independently CN. In another embodiment, RI, 124 and R7 of
formula XIX are
independently OH. In another embodiment, RI, R4 and R7 of formula XIX are
independently alkyl.
In another embodiment, RI, R. and R7 of formula XIX are independently OCH2Ph.
In one
embodiment R2, R5 and R8 of formula XIX are independently H. In another
embodiment, R2, Rs
and R8 of formula XIX are independently 0-alkyl. In another embodiment, R2, R5
and R8 of
CA 3030689 2019-01-18
formula XIX are independently halogen. In another embodiment, R2, R5 and R8of
formula XIX are
independently CN. In another embodiment, R2, R5 and 128 of formula XIX are
independently OH.
In another embodiment, R2, R5 and R8 of formula XIX are independently alkyl.
In another
embodiment, R2, R5 and R8 of formula XIX are independently OCH2Ph. In another
embodiment,
.. R5, R2 and R8 of formula XIX are H, R4 is 4-N(Me)2, R1 is OCH3, m is 3 and
112 is OCH3. In
another embodiment, Rs, R2, R7 and R8 of formula XIX are H, R4 is 4-Br, R1 is
OCH3, and m is 3.
In another embodiment W is S02. In another embodiment W is CO. In another
embodiment W is
C=S. In another embodiment W is S=O. Non limiting examples of compounds of
formula XIX are
selected from: (2-(4-
(dimethylamino)pheny1)- 1-((4-methoxyphenyl)sul fony1)-1H-imidazol-4-
yl)(3,4,5-trimethoxyphenyl)methanone (11gaa); (2-(4-bromopheny1)-1-
(phenylsulfony1)-1H -
imidazol-4-y1)(3,4,5-trimethoxyphenyOmethanone (111a), (4-fluorophenyl)(2-(4-
methoxypheny1)-
1 -(phenylsulfony1)-1H-imidazol-4-yOmethanone (Itch), (2-(4-chloropheny1)-1-
(phenyl sulfon y1)-
1H-i midazol-4-y1)(4-fl uorophenypmethanone (11fb),
(4-fl uorophenyl)(2-pheny1-1-
(phenylsul fony1)-1H-imidazol-4-yOmeth an one (11 af), (4-fluorophenyl)( I -
(phenyl su lfon yI)-2-(p-
tolyI)-1H-imidazol-4-yl)methanone (11db), (2-(4-(dimethylamino)pheny1)-1-
(phenylsulfony1)- I H-
i midazol-411)(3,4,5-trimethox yp hen yl)methanone (11ga),
(2-(4-(dimethylam ino)phenyl )- 1
(phenylsulfony1)- 1H-imidazo1-4-y1)(4-fluorophenypmethanone (11gb), (2-(3,4-
dimethoxypheny1)-
1-(phenylsulfony1)- 1H-imidazol-4-y1)(3,4,5-trimethoxyphenyl)methanone
(11ha), (2-(4-
(benzyloxy)pheny1)- 1-(phenylsulfony1)-1H-imidazol-4-y1)(4-
fluorophenypmethanone (11jb), (2-(4-
(dimethylarnino)pheny1)-1-((4-methox yphenyl)sulfony1)- 1H-imidazol-4-y1)(4-
fluorophenypmethanone (12gba).
[00138] In another embodiment a compound of formula XIX is represented by the
structure of
formula llcb:
0
Me0 ,N di
F
0=S=0
110 ( 1 lcb).
[00139] In another embodiment a compound of formula XIX is represented by the
structure of
formula lift:
71
CA 3030689 2019-01-18
0
110
o=6=o
(11119).
[00140] In one embodiment, this invention is directed to a compound of formula
XX:
0
OMe
R4 ,
NH OMe
OMe
(XX)
wherein
R4 is H, 0-alkyl, I, Br, Cl, F, alkyl, haloalkyl, aminoalkyl, -(CH2),NHCH3, -
(CH2)1NH2, -
(CH2)1N(CH3)2, OCH2Ph, OH, CN, NO1 -NHCO-alkyl, COOH, C(0)0-alkyl or C(0)H;
and
i is an integer between 0-5;
or its pharmaceutically acceptable salt, hydrate, polymorph, metabolite,
tautomer or isomer.
[00141] In one embodiment, R4 of compound of formula VC is H. In another
embodiment, R4 of
compound of formula 30C is halogen. In another embodiment, R. is F. In another
embodiment, R4
is Cl. In another embodiment 1124 is Br. In another embodiment R4 is I. In
another embodiment, R4 is
alkyl. In another embodiment, 114 is methyl. Non limiting examples of
compounds of formula XX
are selected from: (2-pheny1-1H-imidazol-4-y1)(3,4,5-
trimethoxyphenyl)methanone (12aa), (2-(4-
fluoropheny1)-1H-imidazol-4-y1)(3,4,5-trimethoxyphenyl)methanone (12ba),
(2-(4-
methoxypheny1)-1H-imidazol-4-y1)(3,4,5-trimethoxyphenypmethanone (12ca), (2-(p-
toly1)- 1 H-
imidazol-4-y1)(3,4,5-trimethoxyphenyl)methanone (12da), (2-(4-chloropheny1)-1H-
imidazol-4-
y1)(3,4,5-trimethoxyphenyl)methanone (12fa),
(2-(4-(di methyl amino)pheny1)-1H-imi dazol-4-
yl)(3,4,5-trirriethox yphenyl)methanone (12ga), (2-(2-
(trifluoromethyl)pheny1)- I H-imidazol -4-
yl)(3,4,5-trimethox yph eny Dmethan one (121a), (2-(4-(be,nzylox y)phen yl)-1H-
im id azol-4-y1)(3,4,5-
trimethoxyphenyl)methanone (12ja), (2-(4-
hydrox ypheny1)-1H-imidazol-4-y1)(3,4,5-
trimethoxyphenyemethanone (12ka), (2-(4-
bromophen y1)-1H-im id azol-4-y1)(3,4,5-
trimethoxyphenyl)methanone (121a), (2-(4-(trifluoromethyl)pheny1)-1H-imidazol-
4-y1)(3,4,5-
trimethoxyphenyemethanone (12pa).
72
CA 3030689 2019-01-18
[00142] In another embodiment a compound of formula XX is represented by the
structure of
formula 12da:
0
0
HN
-N o
= 0 I
H30 (12da).
[00143] In another embodiment a compound of formula XX is represented by the
structure of
formula 12fa:
0
HN
-N
=
411-11 0
CI (12fa).
[00144] In one embodiment, this invention is directed to a compound of formula
X.XI:
0
__________________________ <
\
(Ri)m
(XXI)
wherein
A is indoly1;
Q is NH, 0 or S;
IS .. R1 and R2 are independently H, 0-alkyl, I, Br, Cl, F, alkyl, haloalkyl,
aminoalkyl, -(CH2)1NHCH3, -
(CH2),NH2, -(CH2)1N(CH3)2, OCH2Ph, OH, CN, NO2, -NICO-alkyl, COOH, C(0)0-alkyl
or
C(0)H; and
wherein said A is optionally substituted by substituted or unsubstituted 0-
alkyl, 0-haloalkyl, F,
Cl, Br, I, haloalkyl, CF3, CN, -CH2CN, NH2, hydroxyl, -(CH2);NHCH3, -
(CH2)1NH2, -
(CH2)1N(CH3)2, -0C(0)CF3, substituted or unsubstituted -S02-aryl, substituted
or unsubstituted
73
=
CA 3030689 2019-01-18
C1-05 linear or branched alkyl, substituted or unsubstituted haloalkyl,
substituted or unsubstituted
alkylamino, substituted or unsubstituted aminoalkyl, -OCH2Ph, substituted or
unsubstituted -
NHCO-alkyl, COOH, substituted or unsubstituted -C(0)Ph, substituted or
unsubstituted C(0)0-
alkyl, C(0)H, -C(0)NH2, NO2 or combination thereof;
i is an integer between 0-5; and
m is an integer between 1-4;
or its pharmaceutically acceptable salt, hydrate, polymorph, metabolite,
tautomer or isomer..
[00145] In one embodiment, RI of compound of formula XXI is OCH3; m is 3 and
R2 is
hydrogen. In another embodiment, R1 is F; m is 1 and R2 is hydrogen. In one
embodiment, Q of
formula XXI is 0. In another embodiment Q of formula XXI is NH. In another
embodiment, Q of
formula XXI is S.
[00146] In one embodiment, A ring of compound of formula XXI is substituted 5-
indolyl. In
another embodiment the substitution is ¨(C=0)-Aryl. In another embodiment, the
aryl is 3,4,5-
(OCH3)3-Ph.
[00147] In another embodiment, A ring of compound of formula XXI is 3-indolyl.
In another
embodiment, A ring of compound of formula XXI is 5-indolyl. In another
embodiment, A ring of
compound of formula XXI is 2-indolyl. Non limiting examples of compounds of
formula XXI are
selected from: (5-(4-(3,4,5-trimethoxybenzoy1)-1H-imidazol-2-y1)-1H-indol-2-
y1)(3,4,5-
trimethoxyphenyl)methanone (15xaa); (1-
(phenylsulfony1)-2-(1-(phenylsulfonyl)-2-(3,4,5-
trimethoxybenzoy1)-1H-indo1-5-y1)-1H-imidazol-4-y1)(3,4,5-
trimethoxyphenyOmethanone (16xaa);
2-(1H-indo1-3-y1)-1H-imidazol-4-y1)(3,4,5-trimethoxyphenyl)methanone (17ya);
(2-(1H-indo1-2-
yl)thiazol-4-y1)(3,4,5-trimethox yphenyl)methanone (62a);
and (2-(1H-indo1-5-ypthiazol-4-
y1)(3,4,5-trimethoxyphenyl)methanone (66a).
[00148] In one embodiment, this invention is directed to a compound of formula
XXIa:
74
CA 3030689 2019-01-18
0
=
ndo e R2
(R
R8
(R7)cl
(XXIa)
wherein
W is C=0, C=S, SO2, S=0;
A is indolyl;
R1 and R2 are independently H, 0-alkyl, I, Br, Cl, F, alkyl, haloalkyl,
aminoalkyl, -(CH2);N1-1CH3, -
(CH2)1NH2, -(CH2)1N(C113)2, OCH2Ph, OH, CN, NO2, -NHCO-alkyl, COOH, C(0)0-
alkyl or
C(0)H;
R7 and Rs are independently H, 0-alkyl, I, Br, Cl, F, alkyl, haloalkyl,
aminoalkyl, -(CH2)iNHCH3, -
(CH2);NH2, -(CH2)1N(CF13)2, OCH2Ph, OH, CN, NO2, -NHCO-alkyl, COOH, C(0)0-
alkyl or
C(0)H;
wherein said A is optionally substituted by substituted or unsubstituted 0-
alkyl, 0-haloalkyl, F,
Cl, Br, I, haloalkyl, CF3, CN, -CH2CN, NH2, hydroxyl, -(CH2)1NHCI-13, -(CH2)N1-
12,
(CH2)1N(CH3)2, -0C(0)CF3, substituted or unsubstituted -S02-aryl, substituted
or unsubstituted
C1-C3 linear or branched alkyl, substituted or unsubstituted haloalkyl,
substituted or unsubstituted
alkylamino, substituted or unsubstituted aminoalkyl, -OCH2Ph, substituted or
unsubstituted -
NHCO-alkyl, COOH, substituted or unsubstituted -C(0)Ph, substituted or
unsubstituted C(0)0-
alkyl, C(0)H, -C(0)NH2, NO2 or combination thereof;
i is an integer between 0-5; and
m is an integer between 1-4;
q is an integer between 1-4;
or its pharmaceutically acceptable salt, hydrate, polymorph, metabolite,
tautomer or isomer.
CA 3030689 2019-01-18
[00149] In one embodiment, R1 of compound of formula XXIa is OCH3; m is 3 and
R2 is
hydrogen. In another embodiment, III is F; m is 1 and R2 is hydrogen. In
another embodiment, A
ring of compound of formula XXIa is substituted 5-indolyl. In another
embodiment, A ring of
compound of formula XXIa is 3-indolyl. Non limiting examples of compounds of
formula XXIa
are selected from: (1:(phenylsulfony1)-2-0-(phenylsulfonyl)-2-(3,4,5-
trimethoxybenzoy1)- 1H -
indo1-5-y1)-1H-imidazol-4-y1)(3,4,5-trimethoxyphenypmethanone (16xaa); (1-
(phenylsulfony1)-2-
( I -(phenylsulfony1)-1H-indo1-3-y1)- I H-imidazol-4-y1)(3,4,5-
trimethoxyphenypmethanone (17yaa).
[00150] In one embodiment, this invention is directed to a compound of formula
XXII:
OCH3
0
=
OCH3
dole
NH OCH3
(XXII)
wherein
A is indolyl;
wherein said A is optionally substituted by substituted or unsubstituted 0-
alkyl, 0-haloalkyl, F,
Cl, Br, I, haloalkyl, CF3, CN, -CH2CN, NH2, hydroxyl, -(CH2)1NHCH3, -(CH2)1141-
12, -
(CH2);N(CH3)2, -0C(0)CF3, substituted or unsubstituted -S02-aryl, substituted
or unsubstituted
C1-05 linear or branched alkyl, substituted or unsubstituted haloalkyl,
substituted or unsubstituted
alkyl amino, substituted or unsubstituted aminoalkyl, -OCH2Ph, substituted or
unsubstituted
NHCO-alkyl, COOH, substituted or unsubstituted -C(0)Ph, substituted or
unsubstituted C(0)0-
alkyl, C(0)H, -C(0)NH2, NO2 or combination thereof;
i is an integer between 0-5;
or its pharmaceutically acceptable salt, hydrate, polymorph, metabolite,
tautomer or isomer.
[00151] In one embodiment, A ring of compound of formula XXII is substituted 5-
indolyl. In
another embodiment the substitution is ¨(C=0)-Aryl. In another embodiment, the
aryl is 3,4,5-
(OCH3)3-Ph.
[00152] In another embodiment, A ring of compound of formula XXII is 3-
indolyl. Non limiting
examples of compounds of formula XXII are selected from: (5-(4-(3,4,5-
trimethoxybenzoy1)-1H-
76
CA 3030689 2019-01-18
imidazol-2-y1)-1H-indo1-2-y1)(3,4,5-trimethoxypheny1)methanone (15xaa); (2-( H-
indo1-3-y1)- I H-
irnidazol -4-yI)(3,4,5-tri methoxyphenyl)methanone (17ya),
[00153] In another embodiment a compound of formula XXI or XXII is represented
by the
structure of formula 17ya:
0
OMe
HN
HN
OMe
=
OMe (17ya).
[00154] In one embodiment, Q of compound of formula XII is 1-1 and P is = .
In
(a
another embodiment, P of compound of formula XII is H and Q is R2 .
In another
embodiment, P of compound of formula XII is R2 and Q
is S02-Ph. In one
41) (RI).
R
embodiment. Q of compound of formula XII is H and P is 2 wherein W is
C=0.
In another embodiment W of compound of formula XII, XVIII, xix, or XXIa is
C=O. In another
embodiment, W of compound of formula XII, XVIII, XIX, or )(Ma is SO2. In
another
embodiment, W of compound of formula XII, XVIII, XIX, or XXIa is C=S. In
another
embodiment, W of compound of formula XII, XVIII, XIX, or XXIa is S=0.
[00155] In one embodiment, Z of compound of formula XIII is oxygen. In another
embodiment,
Z of compound of formula XIII is sulfur.
[00156] In one embodiment, R5 of compound of formula XH-XVI, XVIII, or XIX is
hydrogen, n
is 1 and R4 is in the para position.
77
CA 3030689 2019-01-18
[00157] In one embodiment, R4 of compound of formula XII-XX is alkyl. In
another
embodiment, R4 of compound of formula XII-XX is H. In another embodiment, R4
of compound of
formula XII-XX is methyl (CH3). In another embodiment, R4 of compound of
formula XII-XX is
0-alkyl. In another embodiment, R4 of compound of formula XII-XX is OCH3. In
another
embodiment, Ri of compound of formula WAX is I. In another embodiment, R4 of
compound of
formula XII-XX is Br. In another embodiment, R4 of compound of formula XII-XX
is F. In
another embodiment, R4 of compound of formula XII-MC is Cl. In another
embodiment, R4 of
compound of formula XII-XX is N(Me)2. In another embodiment, R4 of compound of
formula
XII-XX is OBn. In another embodiment, R4 of compound of formula XII-XX is OH.
In another
embodiment, R4 of compound of formula XII-XX is CF3.
[00158] In one embodiment, R2 of compound of formula XII, XIII, XIV, XIVa,
XVII, XIX,
XXI or XXIa is hydrogen; R1 is OCH3 and m is 3. In another embodiment, R2 of
compound of
formula XII, XIII, XIV, XTVa, XVII, XIX, XXI or XXIa is hydrogen; m is 1 and
R1 is in the para
position. In another embodiment, R2 of compound of formula XII, XIII, XIV,
XIVa, XVII, XIX,
XXI or XXIa is hydrogen; m is 1 and R1 is I. In another embodiment, R2 of
compound of formula
XII, XIII, XIV, XIVa, XVII, XIX, XXI or XXIa is hydrogen; m is 1 and R/ is Br.
In another
embodiment, R2 of compound of formula XII, XJU, XIV, XIVa, XVII, XIX, XXI or
XXIa is
hydrogen; m is 1 and R1 is F. In another embodiment, R2 of compound of formula
XII, XIII, XIV,
XIVa, XVII, XIX, XXI or XXIa is hydrogen; m is I and R1 is Cl. In another
embodiment, R1 of
compound of formula XII, XIII, XIV, XIVa, XVII, XIX, XXI or XXIa is I. In
another
embodiment, R1 of compound of formula XII, XHI, XIV, XIVa, XVII, XIX, XXI or
XXIa is Br..
In another embodiment, R1 of compound of formula XII, XM, XIV, XIVa, XVII,
XIX, XXI or
XXIa is Cl. In another embodiment, R1 of compound of formula XII, XIII, XIV,
XIVa, XVII,
XIX, XXI or XXIa is F.
w (RI).
[00159] In one embodiment Q of compound of formula XII is H and P is R2
wherein W is C=0. Non-limiting examples of compounds of formula XII-XVII and
XX-XXII are
selected from (2-pheny1-111-imidazol-4-y1)(3,4,5-trimethoxyphenyl)methanone
(12aa); (4-
78
CA 3030689 2019-01-18
methoxypheny1)(2-pheny1-1H-imidazol-4-yOmethanone (12ab); (3-methoxyphenyl)(2-
pheny1-1H-
.irnidazol -4-yl)methanone (12ac); (3,5-dimethox yphenyl)(2-phTlyl-1H-imidazol-
4-yl)methanone
(12ad); (3,4-dimethoxyphenyl)(2-pheny1-1H-imidazol-4-yl)methanone (12ae); (4-
fluorophenyl)(2-
pheny1-1H-imidazol-4-yOmethanone (12a1); (3-
fluorophenyl)(2:pheny1-1H-imidazol-4-
yl)methanone (12ag); (2-phenyl- I H-imidazol-4-y1)(,p-tolypmethanone (12ah);
(2-phenyl -1 H-
imidazol-4-y1)(m-toly1)methanone (12a1); (2-(4-
fluoropheny1)-1H-imidazol-4-y1)(3,4,5-
trimethoxyphenyl)methanone (12ba); (2-(4-
methoxypheny1)-1H-imidazol-4-y1)(3,4,5-
trimethoxyphenyOmethanone (12ca); (4-
fluorophenyl)(2-(4-methoxypheny1)-1H-imidazol -4-
yflmethanone (12cb); (2-(p-toly1)-1H-imidazol-4-y1)(3,4,5-
trimethoxyphenyl)methanone (12da);
(4-fluorophenyl)(2-(p-toly1)-1H-imidazol-4-yOmethanone (12db); (4-
fluorophenyl)(2-(p-toly1)-1H-
imidazol-4-yl)metharione hydrochloride (12db-HC1); (4-hydrox y-3,5-
dimethoxyphenyl)(2-(p-
toly1)-1H-imidazol-4-yOmethanone (12dc); (3,4,5-trimethoxyphenyl)(2-(3,4,5-
trimethoxypheny1)-
1H-imidazol-4-y1)methanone (12e,a); (4-fluorophenyl)(2-(3,4,5-
trimethoxypheny1)-1H-imidazol-4-
yOmethanone (12eb); (2-(4-ehloropheny1)-1H-imidazol-4-y1)(3,4,5-
trimethoxyphenyl)methanone
(12fa); (2-(4-chloropheny1)-1H-imidazol-4-y1)(4-fluorophenyl)methanone (12th);
(2-(4-
chloropheny1)-1H-imidazol-4-y1)(4-hydroxy-3,5-dimethoxyphenypmethanone
(12fc); (2-(4-
(dimethylaniino)pheny1)-1H-imidazol-4-y1)(3,4,5-trimethoxyphenyl)methanone
(12ga); (2-(4-
(dimethylarnino)pheny1)-1H-imidazol-4-y1)(4-fluorophenyl)methanone (12gb);
(243,4-
d imethoxypheny1)-1H-imi dazol-4-y1)(3,4,5-trimethoxyphenypmethanone
(12ha); (2-(3,4-
dimethoxypheny1)-1H-imidazol-4-y1)(4-fluorophenypmethanone (12hb);
(2-(2-
(trifluoromethyl)pheny1)-1H-irnidazol-4-y1)(3,4,5-trimethoxyphenyl)methanone
(121a); (4-
fl uorophenyl)(2-(2-(tri fl uoromethyl)pheny1)-111-imidazol-4-yOmethanone
(121b); (2-(4-
(benzyloxy)pheny1)-1H-imidazol-4-y1)(3,4,5-trimethoxyphenyl)nnethanone
(12ja); (2-(4-
(benzyloxy)pheny1)-1H-imidazol-4-y1)(4-fluorophenypmethanone (12jb); (2-(4-
hydrox ypheny1)-
1H-imidazol-4-y1)(3,4,5-trimethoxyphenyl)methanone (12ka); (2-
(4-(h ydroxyphen y1)- 1 H-
i midazol-4-y1)(4-fluorophenyl)methanone (12kb); (2-(4-bromopheny1)-1H-
imidazol-4-y1)(3,4,5-
trimethoxyphenyl)methanone (121a); (2-(4-(trifluoromethyl)pheny1)-1H-imidazol-
4-y1)(3,4,5-
trimethoxyphenyOmethanone (12pa); (3,4,5-trihydrox yphenyl)(2-(3,4,5-trihydrox
yphen y1)- 1 H-
imidazol-4-yOmethanone (13ea); (2-(4-
ch loropheny1)-1H-i midazol-4-y1)(3,4,5-
trihydroxyphenyl)methanone (13ra); and 2-(3,4-dihydroxypheny1)-1H-imidazol-4-
y1)(3,4,5-
trihydrox yphenypmethanone (13ha).
79
=
CA 3030689 2019-01-18
411 (RI)m
=
[00160] In one embodiment, P of compound of formula XII is R2
and Q is 802-
Ph. Non-limiting examples of compound of formula XII wherein P of compound of
formula XII is
(RI).
R2
and Q is 802-Ph are selected from (4-methoxyphenyl)(2-pheny1-1-
(phenyl s ulfony1)-1H-imidazol-4-y1)methanone (nab);
(3-methoxyphenyl)(2-pheny1-1-
(phenyl sulfony1)-1H-imidazol-4-yOmethanone (11ac); (2-pheny1-1-
(phenylsulfony1)-1H-irnidazol-
4-y1)(p-tolyOmethanone (11ah); (4-fl uorophenyl)(2-pheny1-1-(phenyl sul fon
y1)-1H-imidazol-4-
yOmethanone (11af); (3-fluorophenyl)(2-phenyl-1-(phenylsulfony1)-1H-iinidazol-
4-yOmethanone
(hag); (4 -
fluorophenyl)(2-(4-methox ypheny1)-1-(phenylsulfony1)-1H-imidazol-4-
y1)methanone
(11cb); (1-
(phenylsulfony1)-2-(p-toly1)-1H-imidazol-4-y1)(3,4,5-trimethox
yphenyflmethanone
.. (11da) ; (4 -fluorophenyl )(1-(phenylsulfony1)-2-(p-tol y1)-1H-imidazol-4-
y1)methanone (Ildb); (1-
(phenylsulfony1)-2-(3,4,5-trimethox ypheny1)-1H-imidazol-4-y1)(3,4,5-tri
methoxyphenyflmethanone
(ilea); (4-
fluorophenyl)( -(phenylsulfony1)-2-(3,4,5-trimethox ypheny1)-1H-i mi dazol-4-
yl)methanone (11cb); (2-(4-
chloropheny1)-1-(phenylsulfony1)-1H-imidazol-4-y1)(4-
fluorophenyl)methanone (1111)); (2-(4-(dimethylamino)pheny1)-1-
(phenylsulfony1)-1H-imidazol-4-
yl)(3,4,5-trimethox yphenypmethanone (11ga); (2-(4-(dimethylami no)pheny1)-1-
(phen yl sulfon yI)-
I H-iinidazol-4-y1)(4-fluorophenyOmethanone (1
lgb); (2-(3,4-dimethoxypheny1)-1-
(phenylsulfony1)-1H-imidazol-4-y1)(3,4,5-trimethoxyphenyl)methanone
(11ha); .. (243,4-
di methox ypheny1)-1-(phenylsulfony1)-1H-imidazol-4-y1)(4-
fluorophenyl)methanone (11hb); (1-
.
(phenylsul fony1)-2-(2-(trifluoromethyl)pheny1)-1H-i midazol-4-y1)(3,4 ,5-
trimethox yphenyl)methanone (ilia); (1-(phenylsulfony1)-2-
(2-(trifluoromethyl)pheny1)-1H-
imidazol-4-y1)(4-fluorophenypmethanone (llib); and
(2 -(4-(benzylox y)pheny1)- I -
(phenylsulfony1)-1H-imidazol-4-y1)(4-fluorophenyl)methanone (11jb); (2-(4-
bromopheny1)-1-
(phenyl s ulfony1)-1H-imidazol-4-y1)(3,4,5 -trimethoxyphen yl )methanone
(111a); (1-
(phenyls ulfony1)-2-(4-(tri fluoromethyppheny1)-1H-i midazol-4-y1)(3,4,5-
trimethoxyphenyl)methanone (11pa).
[00161] In one embodiment, R4 and R5 of compounds of formula are
hydrogens. Non-
limiting examples of compounds of formula XLII-XVI wherein R4 and R5 are
hydrogens are
=
CA 3030689 2019-01-18
selected from (2-phenyl-1H-imidazol-4-y1)(3,4,5-trimethoxyphenyl)methanone
(12aa); (4-
methoxyphenyl)(2-pheny1-1H-i midazol-4-yOmethanone (12ab); (3-methoxyphenyl)(2-
phenyl-1H-
imidazol-4-yl)methanone (12ac); (3,5-di methoxyphen yl )(2-pheny1-1H-imidazol-
4-yl)methanone
(12ad); (3,4-dimethoxyphenyl)(2-pheny1-1/1-imidazol-4-y1)methanone (12ae); (4-
fluorophenyl)(2-
phenyl-1H-imidazol-4-yl)methanone (12af); (3-fluorophenyl)(2-pheny1-1H-
imidazol-4-
yOmethanone (12ag); (2-phenyl-1H-imidazol-4-y1)(p-tolyl)methanone (12ah); and
(2-phenyl-1H-
imidazol-4-y1)(m-tolypmethanone (12a1).
w (111)m
R2
[00162) In one embodiment, P of compound of formula XTI is H and Q is . In
another embodiment W is C=0. In another embodiment, W of compound of formula
XVIII is
C=0. Non-limiting examples of compound of formula XVIII wherein W is C=0 are
selected from
(4-methoxyphenyl)(2-pheny1-1H-imidazol-1-yOmethanone (12aba) and (2-phenyl-I H-
imidazol-1-
yl)(3,4,5-trimethoxyphenypmethanone (12aaa).
[00163] In another embodiment, W of compound of formula XVIII is SO2. Non-
limiting
examples of compound of formula XVIII wherein W is SO2 are selected from 2-
phenyl-1-
(phenylsulfony1)-1H-imidazole (10a); 2-(4-nitropheny1)-1-(phenylsulfony1)-1H-
imidazole (10x)
and 2-(4-(benzyloxy)pheny1)-1-(phenylsulfony1)-1H-imidazole (10j).
[00164] As used
herein, "single-, fused- or multiple-ring, aryl or (hetero)cydlic ring
systems" can be any such ring, including but not limited to phenyl, biphenyl,
triphenyl, naphthyl,
cycloalkyl, cycloalkenyl, cyclodienyl, fluorene, adamantane, etc.
[00165) "Saturated
or unsaturated N-heterocycles" can be any such N-containing
heterocycle, including but not limited to aza- and diaza-cycloalkyls such as
aziridinyl, azetidinyl,
diazatidinyl, pyrrolidinyl, piperidinyl, piperazinyl, and azocanyl, pyrrolyl,
pyrazolyl,
pyridinyl, pyrimidinyl, pyrazinyl, pyridazinyl, triazinyl, tetrazinyl,
pyrrolizinyl, indolyl,
quinolinyl, isoquinolinyl, benzimidazolyl, indazolyl, quinolizinyl,
cinnolinyl, quinololinyl,
phthalazinyl, naphthyridinyl, quinoxalinyl, etc.
[00166]
"Saturated or unsaturated 0-Heterocycles" can be any such 0-containing
heterocycle including but not limited to oxiranyl, oxetanyl,
tetrahydrofuranyl, tetrahydropyranyl,
dioxanyl, furanyl, pyrylium, benzofuranyl, benzodioxolyl, etc.
81
CA 3030689 2019-01-18
[00167] "Saturated or unsaturated S-heterocycles" can be any such S-
containing
heterocycle, including but not limited to thiranyl, thietanyl,
tetrahydrothiophene-yl, dithiolanyl,
tetrahydrothiopyranyl, thiophene-yl, thiepinyl, thianaphthenyl, etc.
[00168] "Saturated or unsaturated mixed heterocycles" can be any
heterocycle containing
two or more S-, N-, or 0-heteroatoms, including but not limited to
oxathiolanyl, morpholinyl,
thioxanyl, thiazolyl, isothiazolyl, thiadiazolyl, oxazolyl, isoxazolyl,
oxadiaziolyl, etc.
[001691 As used herein, "aliphatic straight- or branched-chain
hydrocarbon" refers to both
alkylene groups that contain a single carbon and up to a defined upper limit,
as well as alkenyl
groups and alkynyl groups that contain two carbons up to the upper limit,
whether the carbons are
present in a single chain or a branched chain. Unless specifically identified,
a hydrocarbon can
include up to about 30 carbons, or up to about 20 hydrocarbons, or up to about
10 hydrocarbons.
Alkenyl and alkynyl groups can be mono-unsaturated or polyunsaturated. In
another
embodiment, an alkyl includes C1-05 carbons. In another embodiment, an alkyl
includes C1-C8
carbons. In another embodiment, an alkyl includes C1-C10 carbons. In another
embodiment, an
alkyl is a C1-C12 carbons. In another embodiment, an alkyl is a C1-05 carbons.
[00170) As used herein, the term "alkyl" can be any straight- or
branched-chain alkyl
= group containing up to about 30 carbons unless otherwise specified. In
another embodiment, an
alkyl includes C1-05 carbons. In another embodiment, an alkyl includes C1-C8
carbons. In another
embodiment, an alkyl includes C1-C10 carbons. In another embodiment, an alkyl
is a CI-C12
carbons. In another embodiment, an alkyl is a C1-C20 carbons. In another
embodiment, cyclic
alkyl group has 3-8 carbons. In another embodiment, branched alkyl is an alkyl
substituted by
alkyl side chains of 1 to 5 carbons.
[00171] The alkyl group can be a sole substituent or it can be a
component of a larger
substituent, such as in an alkoxy, haloalkyl, arylalkyl, alkylamino,
dialkylamino, alkylamido,
alkylurea, etc. Preferred alkyl groups are methyl, ethyl, and propyl, and thus
halomethyl,
dihalomethyl, trihalomethyl, haloethyl, dihaloethyl, trihaloethyl, halopropyl,
dihalopropyl,
trihalopropyl, methoxy, ethoxy, propoxy, arylmethyl, arylethyl, arylpropyl,
methylamino,
ethylamino, propylamino, dimethylamino, diethylamino, methylamido, acetarnido,
propylamido,
halomethylamido, haloethylamido, halopropylamido, methyl-urea, ethyl-urea,
propyl-urea, etc.
82
CA 3030689 2019-01-18
[00172] As used herein, the term "aryl" refers to any aromatic ring
that is directly bonded
to another group. The aryl group can be a sole substituent, or the aryl group
can be a component
of a larger substituent, such as in an arylalkyl, arylamino, arylamido, etc.
Exemplary aryl groups
include, without limitation, phenyl, tolyl, xylyl, furanyl, naphthyl,
pyridinyl, pyrimidinyl,
pyridazinyl, pyrazinyl, .triazinyl, thiazolyl, oxazolyl, isooxazolyl,
pyrazolyl, imidazolyl,
thiophene-yl, pyrrolyl, phenylmethyl, phenylethyl, phenylamino, phenylarnido,
etc.
[00173] As used herein, the term "arninoalkyl" refers to an amine group
substituted by an
alkyl group as defined above. Aminoalkyl refers to monoalkylamine,
dialkylarnine or triallcylamine.
Nonlimiting examples of aminoalkyl groups are -N(Me)2, -NHMe, -NH3.
[00174] A "haloalkyl" group refers, in another embodiment, to an alkyl
group as defined
above, which is substituted by one or more halogen atoms, e.g. by F, Cl, Br or
I. Nonlimiting
examples of haloalkyl groups are CF3, CF2CF3, CH2CF3.
[00175] In one embodiment, this invention provides a compound of this
invention or its
isomer, metabolite, pharmaceutically acceptable salt, pharmaceutical product,
tautomer, hydrate, N-
oxide, polymorph, or crystal or combinations thereof. In one embodiment, this
invention provides an
isomer of the compound of this invention. In another embodiment, this
invention provides a
metabolite of the compound of this invention. In another embodiment, this
invention provides a
pharmaceutically acceptable salt of the compound of this invention. In another
embodiment, this
invention provides a pharmaceutical product of the compound of this invention.
In another
embodiment, this invention provides a tautomer of the compound of this
invention. In another
embodiment, this invention provides a hydrate of the compound of this
invention. In another
embodiment, this invention provides an N-oxide of the compound of this
invention. In another
embodiment, this invention provides a polymorph of the compound of this
invention. In another
embodiment, this invention provides a crystal of the compound of this
invention. In another
embodiment, this invention provides composition comprising a compound of this
invention, as
described herein, or, in another embodiment, a combination of an isomer,
metabolite,
pharmaceutically acceptable salt, pharmaceutical product, tautomer,hydrate, N-
oxide, polymorph, or
crystal of the compound of this invention.
[00176] In one embodiment, the term "isomer" includes, but is not
limited to, optical isomers
and analogs, structural isomers and analogs, conformational isomers and
analogs, and the like.
83
CA 3030689 2019-01-18
[00177] In one embodiment, the compounds of this invention are the pure
(E)-isomers. In
another embodiment, the compounds of this invention are the pure (Z)-isomers.
In another
embodiment, the compounds of this invention are a mixture of the (E) and the
(Z) isomers. In one
embodiment, the compounds of this invention are the pure (R)-isomers. In
another embodiment, the
compounds of this invention are the pure (S)-isomers. In another embodiment,
the compounds of
this invention are a mixture of the (R) and the (S') isomers.
[00178] The compounds of the present invention can also be present in
the form of a racemic
mixture, containing substantially equivalent amounts of stereoisomers. In
another embodiment, the
compounds of the present invention can be prepared or otherwise isolated,
using known procedures,
to obtain a stereoisomer substantially free of its corresponding stere,oisomer
(i.e., substantially pure).
By substantially pure, it is intended that a stereoisomer is at least about
95% pure, more preferably at
least about 98% pure, most preferably at least about 99% pure.
[00179] Compounds of the present invention can also be in the form of a
hydrate, which
means that the compound further includes a stoichiometric or non-
stoichiometric amount of water
bound by non-covalent intermolecular forces.
[00180] Compounds of the present invention may exist in the form of one
or more of the
possible tautomers and depending on the particular conditions it may be
possible to separate
some or all of the tautomers into individual and distinct entities. It is to
be understood that all of
the possible tautomers, including all additional enol and keto tautomers
and/or isomers are hereby
covered. For example the following tautomers, but not limited to these, are
included.
Tautomerization of the imidazole ring
H N _ N-Y
[00181] The invention includes "pharmaceutically acceptable salts" of
the compounds of
this invention, which may be produced, by reaction of a compound of this
invention with an acid
or base. Certain compounds, particularly those possessing acid or basic
groups, can also be in the
form of a salt, preferably a pharmaceutically acceptable salt. The term
"pharmaceutically
acceptable salt" refers to those salts that retain the biological
effectiveness and properties of the
free bases or free acids, which are not biologically or otherwise undesirable.
The salts are formed
with inorganic acids such as hydrochloric acid, hydrobromic acid, sulfuric
acid, nitric acid,
84
CA 3030689 2019-01-18
phosphoric acid and the like, and organic acids such as acetic acid, propionic
acid, glycolic acid,
pyruvic acid, oxylic acid, maleic acid, malonic acid, succinic acid, fumaric
acid, tartaric acid,
citric acid, benzoic acid, cinnamic acid, mandelic acid, methanesulfonic acid,
ethanesulfonic acid,
p-toluenesulfonic acid, salicylic acid, N-acetylcysteine and the like. Other
salts are known to
those of skill in the art and can readily be adapted for use in accordance
with the present
invention.
[00182]
Suitable pharmaceutically-acceptable salts of amines of compounds the
compounds of this invention may be prepared from an inorganic acid or from an
organic acid. In
one embodiment, examples of inorganic salts of amines are bisulfates, borates,
bromides,
chlorides, hemisulfates, hydrobromates,
hydrochlorates, 2-hydrox yethylsulfonates
(hydroxyethanesulfonates), iodates, iodides, isothionates, nitrates,
persulfates, phosphate,
sulfates, sulfamates, sulfanilates, sulfonic acids (alkylsulfonates,
arylsulfonates, halogen
substituted alkylsulfonates, halogen substituted arylsulfonates), sulfonates
and thiocyanates.
[00183] In one
embodiment, examples of organic salts of amines may be selected from
aliphatic, cycloaliphatic, aromatic, araliphatic, heterocyclic,scarboxylic and
sulfonic classes of
organic acids, examples of which are acetates, arginines, aspartates,
ascorbates, adipates,
anthranilates, algenates, alkane carboxylates, substituted alkane
carboxylates, alginates,
benzenesulfonates, benzoates, bisulfates, butyrates, bicarbonates,
bitartrates, citrates,
camphorates, camphorsulfonates, cyclohexylsulfamates, cyclopentanepropionates,
calcium
edetates, camsylates, carbonates, clavulanates, cinnamates, dicarboxylates,
digluconates,
dodecylsulfonates, dihydrochlorides, decanoates, enanthuates,
ethanesulfonates, edetates,
edisylates, estolates, esylates, fumarates, formates, fluorides,
galacturonates gluconates,
glutamates, glycolates, glucorate, glucoheptanoates, glycerophosphates,
gluceptates,
glycollylarsani I ates, glutarates, glutamate, heptanoates, hexanoates,
hydroxymaleates,
hydroxycarboxlic acids, hex ylresorci nates, hydrox
ybenzoates, hyd rox yn aph thoates,
hydrofluorates, lactates, lactobionates, laurates, malates, maleates,
methylenebis(beta-
oxynaphttioate), malonates, mandelates, mesylates, methane sulfonates, meth
ylbromides,
methylnitrates, methylsulfonates, monopotassium maleates, mucates,
monocarboxylates,
naphthalenesulfonates, 2-naphthalenesulfonates, nicotinates, nitrates,
napsylates, N-
methylglucarnines, oxalates, octanoates, oleates, pamoates, phenylacetates,
picrates,
phenylbenzoates, pivalates, propionates, phthalates, phenylacetate,
pectinates, phenylpropionates,
CA 3030689 2019-01-18
palmitates, pantothenates, polygalacturates, pyruvates, quinates, salicylates,
succinates, stearates,
sulfanilate, subacetates, tartrates, theophyllineacetates, p-toluenesulfonates
(tosYlates),
trifluoroacetates, terephthalates, tannates, teoclates, trihaloacetates,
triethiodide, tricarboxylates,
undecanoates and valerates.
[00184] In one embodiment, examples of inorganic salts of carboxylic acids
or hydroxyls
may be selected from ammonium, alkali metals to include lithium, sodium,
potassium, cesium; .
alkaline earth metals to include calcium, magnesium, aluminium; zinc, barium,
cholines,
quaternary ammoniums.
[00185] In another embodiment, examples of organic salts of carboxylic
acids or hydroxyl
may be selected from arginine, organic amines to include aliphatic organic
amines, alicyclic
organic amines, aromatic organic amines, benzathines, t-butylamines,
benethamines (N-
benzylphenethylamine), dicyclohexylamines, dimethylamines, diethanolamines,
ethanolamines,
ethylenediamines, hydrabamines, imidazoles, lysines, methyl amines,
meglamines, /V-methyl-D-
glucamines, N,N'-dibenzylethylenediamines, nicotinamides, organic amines,
ornithines,
pyridines, picolies, piperazines, procain, tris(hydroxymethyl)methylamines,
triethylamines,
triethanolamines, trimethylamines, trometharnines and ureas.
[00186] In one embodiment, the salts may be formed by conventional
means, such as by
reacting the free base or free acid form of the product with one or more
equivalents of the
appropriate acid or base in a solvent or medium in which the salt is insoluble
or in a solvent such
.. as water, which is removed in vacuo or by freeze drying or by exchanging
the ions of a existing
salt for another ion or suitable ion-exchange resin.
[00187] In some embodiments, this invention provides a process for the
preparation of the
compounds of this invention. In one embodiment, the aryl-imidazole is prepared
by reacting an
appropriately substituted benzaldehyde with ethylenediamine to construct the
imidazoline ring,
.. followed by oxidation of the imidazoline by an oxidizing agent to the
corresponding irnidazole.
In another embodiment the oxidizing agent is diacetoxyiodobenzene,
bromotrichloromethane and
1,8-diazabicyclo[5.4.0]undec-7-ene (DBU), carbon-02 system or palladium-carbon
system. In
another embodiment, the aryl-imidazole is prepared by reacting an
appropriately substituted
benzaldehyde with ethylene diamine in the presence of iodine and potassium
carbonate in order
to construct the imidazoline ring, followed by oxidation of the imidazoline
ring catalyzed by
86
CA 3030689 2019-01-18
diacetoxyiodobenzene, bromotrichloromethane and 1,8-diazabicyclo[5.4.0]undec-7-
ene (DBU),
carbon-02 system or palladium-carbon system to the corresponding imidazole. In
another
embodiment, the aryl-imidazole is prepared by reacting an appropriately
substituted
benzaldehyde with ethylene diarnine in the presence of iodine and potassium
carbonate in order
to construct the imidazoline ring, followed by oxidation of the imidazoline
ring catalyzed by
diacetoxyiodobenzene to the corresponding imidazole. In another embodiment,
the aryl-
imidazole is prepared by reacting an appropriately substituted benzaldehyde
with ethylene
diamine in the presence of iodine and potassium carbonate-in order to
construct the imidazoline
ring, followed by oxidation of the imidazoline ring catalyzed by
bromotrichloromethane and 1,8-
diazabicyclo[5.4.0]undec-7-ene (DBU) to the corresponding imidazole. In one
embodiment, the
aryl-imidazole is prepared by reacting the appropriate benzaldehyde in ethanol
with oxalaldehyde
and ammonia hydroxide to construct the imidazole ring system.
[00188] In one embodiment an aryl-benzoyl-imidazole compound of this
invention is
prepared by protecting the aryl-imidazole followed by coupling with an
appropriately substituted
benzoyl chloride, followed by removing the protecting group. In another
embodiment, the
protecting group is a phenyl sulfonyl group, phthalimide, di-tert-butyl
dicarbonate (Boc),
fluorenylmethyloxycarbonyl (Fmoc), benzyloxycarbonyl (Cbz), or
monomethoxytrityl (MMT).
In another embodiment, the aryl-imidazole is protected with phenyl sulfonyl to
yield the N-
sulfonyl protected aryl-imidazole. In another embodiment, the protected aryl-
imidazole
compound is prepared by reacting the aryl-imidazole with phenylsulfonyl
chloride and sodium
hydride in THF. In another embodiment, the protected aryl-imidazole is
prepared according to
Figures 7 and 8.
[00189] In one embodiment, the protected aryl-imidazole is coupled with
an appropriately
substituted benzoyl chloride to obtain a protected aryl-benzoyl imidazole. In
another
embodiment, aryl-imidazole is coupled with an appropriately substituted
benzoyl chloride in the
presence of tert-butyl lithium to obtain aryl-phenylsulfonyl (2-ary1-1-
(phenylsulfony1)-1H-
imidazol-4-y1)methanone. In another embodiment, the (2-ary1-1-(phenylsulfony1)-
1H-imidazol-4-
y1)methanone is prepared according to Figures 7 and 8 steps e and c,
respectively.
[00190] In one embodiment, an aryl-benzoyl-imidazole is prepared by
removing the
protecting group of the aryl-benzoyl-imidazole. In another embodiment, the
removal of the
protecting group depends on the protecting group used and can be removed by
known conditions
87
CA 3030689 2019-01-18
which are known in the art. In another embodiment, the phenyl sulfonyl
protecting group is
removed by tetrabutylammonium fluoride in THF. In another embodiment,
phenylsulfonyl is
removed according to Figures 7 and 8.
[00191] In one embodiment, compounds of formula 1, Ia, II, III, V and
XI are prepared
according to Figure 1. In another embodiment, compounds of formula I, Ia, II,
III, V, VI, VII
and XI are prepared according to Figure 2. In another embodiment, compounds of
formula I, Ia,
II, III, V and VI are prepared according to Figure 3. In another embodiment,
compounds of
formula I, Ia, II, III, V and VI are prepared according to Figure 4. In
another embodiment,
compounds of formula I, Ia, II, III, IV, IVa, V, VI and XI are prepared
according to Figure 5.
In another embodiment, compounds of formula I, Ia, II, III, VIII and XI are
prepared according
to Figure 6.
= [00192] In one embodiment, compounds of formula XII and
XVIII are prepared according
to Figure 9. In another embodiment, compounds of formula XII, XIII, XIV, XIVa,
XV, XVI,
XVII, XIX and XX are prepared according to Figure 10. In another embodiment,
compounds of
formula XIVa and XIX are prepared according to Figure 11. In another
embodiment, compounds
of formula I, Ia, IV, IVa, XI, XXI, XXIa and XXII are prepared according to
Figure 12. In
=
another embodiment, compounds of formula I, Ia, IV, IVa, XI, XIb, XXI, XXIa
and XXII are
prepared according to Figure 13. In another embodiment, compounds of formula
I, Ia, II, III, V,
XI, XII, XIII, XIV, XV, XVII, XIX and XX are prepared according to Figure 14.
In another
embodiment, compounds of formula I, Ia, H, IV, IVa, XI and XIc, are prepared
according to
Figure 15.
[00193] In one embodiment, compounds of formula IX and IXa are prepared
according to
Figure 16.
Pharmaceutical composition
[00194] Another aspect of the present invention relates to a pharmaceutical
composition including
a pharmaceutically acceptable carrier and a compound according to the aspects
of the present
invention. The pharmaceutical composition can contain one or more of the above-
identified
compounds of the present invention. Typically, the pharmaceutical composition
of the present
invention will include a compound of the present invention or its
pharmaceutically acceptable salt,
88
CA 3030689 2019-01-18
as well as a pharmaceutically acceptable carrier. The term "pharmaceutically
acceptable carrier"
refers to any suitable adjuvants, carriers, excipients, or stabilizers, and
can be in solid or liquid form
such as, tablets, capsules, powders, solutions, suspensions, or emulsions.
1001951 Typically, the composition will contain from about 0.01 to 99 percent,
preferably from
about 20 to 75 percent of active compound(s), together with the adjuvants,
carriers and/or excipients.
While individual needs may vary, determination of optimal ranges of effective
amounts of each
component is within the skill of the art. Typical dosages comprise about 0.01
to about 100 mg/kg
body wt. The preferred dosages comprise about 0.1 to about 100 mg/kg body wt.
The most preferred
dosages comprise about 1 to about 100 mg/kg body wt. Treatment regimen for the
administration of
the compounds of the present invention can also be determined readily by those
with ordinary skill
in art That is, the frequency of administration and size of the dose can be
established by routine
optimization, preferably while minimizing any side effects.
[00196] The solid unit dosage forms can be of the conventional type. The solid
form can be a
capsule and the like, such as an ordinary gelatin type containing the
compounds of the present
invention and a carrier, for example, lubricants and inert fillers such as,
lactose, sucrose, or
cornstarch. In another embodiment, these compounds are tabulated with
conventional tablet. bases
such as lactose, sucrose, or cornstarch in combination with binders like
acacia, cornstarch, or gelatin,
disintegrating agents, such as cornstarch, potato starch, or alginic acid, and
a lubricant, like stearic
acid or magnesium stearate.
[00197] The tablets, capsules, and the like can also contain a binder such as
gum tragacanth,
acacia, corn starch, or gelatin; excipients such as dicalcium phosphate; a
disintegrating agent such as
corn starch, potato starch, alginic acid; a lubricant such as magnesium
stearate; and a sweetening
agent such as sucrose, lactose, or saccharin. When the dosage unit form is a
capsule, it can contain,
in addition to materials of the above type, a liquid carrier such as a fatty
oil.
[00198] Various other materials may be present as coatings or to modify the
physical form of the
dosage unit. For instance, tablets can be coated with shellac, sugar, or both.
A syrup can contain, in
addition to active ingredient, sucrose as a sweetening agent, methyl and
propy/parabens as
preservatives, a dye, and flavoring such as cherry or orange flavor.
[00199] For oral therapeutic administration, these active compounds can be
incorporated with
excipients and used in the form of tablets, capsules, elixirs, suspensions,
syrups, and the like. Such
89
=
CA 3030689 2019-01-18
compositions and preparations should contain at least 0.1% of active compound.
The percentage of
the compound in these compositions can, of course, be varied and can
conveniently be between
about 2% to about 60% of the weight of the unit. The amount of active compound
in such
therapeutically useful compositions is such that a suitable dosage will be
obtained_ Preferred
compositions according to the present invention are prepared so that an oral
dosage unit contains
between about 1 mg and 800 mg of active compound.
[00200] The active compounds of the present invention may be orally
administered, for example,
with an inert diluent, or with an assimilable edible carrier, or they can be
enclosed in hard or soft
shell capsules, or they can be compressed into tablets, or they can be
incorporated directly with the
food of the diet.
[00201] The pharmaceutical forms suitable for injectable use include sterile
aqueous solutions or
dispersions and sterile powders for the extemporaneous preparation of sterile
injectable solutions or
dispersions. In all cases, the form should be sterile and should be fluid to
the extent that easy
syringability exists. It should be stable under the conditions of manufacture
and storage and should
be preserved against the contaminating action of microorganisms, such as
bacteria and fungi. The
carrier can be a solvent or dispersion medium containing, for example, water,
ethanol, polyol (e.g.,
glycerol, propylene glycol, and liquid polyethylene glycol), suitable mixtures
thereof, and vegetable
oils.
[00202] The compounds or pharmaceutical compositions of the present invention
may also be
.. administered in injectable dosages by solution or suspension of these
materials in a physiologically
acceptable diluent with a pharmaceutical adjuvant, carrier or excipient. Such
adjuvants, carriers
and/or excipients include, but are not limited to, sterile liquids, such as
water and oils, with or
without the addition of a surfactant and other pharmaceutically and
physiologically acceptable
components. Illustrative oils are those of petroleum, animal, vegetable, or
synthetic origin, for
example, peanut oil, soybean oil, or mineral oil. In general, water, saline,
aqueous dextrose and
related sugar solution, and glycols, such as propylene glycol or polyethylene
glycol, are preferred
liquid carriers, particularly for injectable solutions.
[00203] These active compounds may also be administered parenterally.
Solutions or suspensions
of these active compounds can be prepared in water suitably Mixed with a
surfactant such as
.. hydroxypropylcellulose. Dispersions can also be prepared in glycerol,
liquid polyethylene glycols,
=
CA 3030689 2019-01-18
and mixtures thereof in oils. Illustrative oils are those of petroleum,
animal, vegetable, or synthetic
origin, for example, peanut oil, soybean oil, or mineral oil. In general,
water, saline, aqueous
dextrose and related sugar solution, and glycols such as, propylene glycol or
polyethylene glycol, are
preferred liquid carriers, particularly for injectable solutions. Under
ordinary conditions of storage
and use, these preparations contain a preservative to prevent the growth of
microorganisms.
[00204] For use as aerosols, the compounds of the present invention in
solution or suspension may
be packaged in a pressurized aerosol container together with suitable
propellants, for example,
hydrocarbon propellants like propane, butane, or isobutane with conventional
adjuvants. The
materials of the present invention also may be administered in a non-
pressurized form such as in a
nebulizer or atomizer.
[00205] In one embodiment, the compounds of this invention are
administered in
combination with an anti-cancer agent. In one embodiment, the anti-cancer
agent is a monoclonal
antibody. In some embodiments, the monoclonal antibodies are used for
diagnosis, monitoring, or
treatment of cancer. In one embodiment, monoclonal antibodies react against
specific antigens on
.. cancer cells. In one embodiment, the monoclonal antibody acts as a cancer
cell receptor
antagonist. In one embodiment, monoclonal antibodies enhance the patient's
immune response. In
one embodiment, monoclonal antibodies act against cell growth factors, thus
blocking cancer cell
growth. In one embodiment, anti-cancer monoclonal antibodies are conjugated or
linked to anti-
cancer drugs, radioisotopes, other biologic response modifiers, other toxins,
or a combination
thereof. In one embodiment, anti-cancer monoclonal antibodies are conjugated
or linked to a
compound of this invention as described hereinabove.
[00206] Yet another aspect of the present invention relates to a method of
treating cancer that
includes selecting a subject in need of treatment for cancer, and
administering to the subject a
pharmaceutical composition comprising a compound according to the first aspect
of the present
invention and a pharmaceutically acceptable carrier under conditions effective
to treat cancer.
[00207] When administering the compounds of the present invention, they can be
administered
systemically or, alternatively, they can be administered directly to a
specific site where cancer cells
or precancerous cells are present. Thus, administering can be accomplished in
any manner effective
for delivering the compounds or the pharmaceutical compositions to the cancer
cells or precancerous
cells. Exemplary modes of administration include, without limitation,
administering the compounds
91
CA 3030689 2019-01-18
or compositions orally, topically, transdermally, parenterally,
subcutaneously, intravenously,
intramuscularly, intraperitoneally, by intranasal instillation, by
intracavitary or intravesical
instillation, intraocularly, intraarterially, intralesionally, or by
application to mucous membranes,
such as, that of the nose, throat, and bronchial tubes_
Biological Activily
[00208] , In one embodiment, the invention provides compounds and
compositions,
including any embodiment described herein, for use in any of the methods of
this invention. In
one embodiment, use of a compound of this invention or a composition
comprising the same, will
have utility in inhibiting, suppressing, enhancing or stimulating a desired
response in a subject, as
will be understood by one skilled in the art. In another embodiment, thc
compositions may
further comprise additional active ingredients, whose activity is useful for
the particular
application for which the compound of this invention is being administered.
[00209] In one embodiment, this invention is directed to a method of treating,
suppressing,
reducing the severity, reducing the risk of developing or inhibiting cancer
comprising administering
a compound of this invention to a subject suffering from cancer under
conditions effective to treat
the cancer.
[00210] Drug resistance is the major cause of cancer chemotherapy
failure. One major
contributor to multidrug resistance is overexpression of P-glycoprotein (P-
gp). This protein is a
clinically important transporter protein belonging to the ATP-binding cassette
family of cell
membrane transporters. It can pump substrates including anticancer drugs out
of tumor cells
through an ATP-dependent mechanism.
[00211] In one embodiment, this invention provides methods for: a)
treating, suppressing,
reducing the severity, reducing the risk, or inhibiting drug resistant tumors;
b) treating,
suppressing, reducing the severity, reducing the risk, or inhibiting
metastatic cancer; c) treating,
suppressing, reducing the severity, reducing the risk, or inhibiting drug
resistant cancer; d)
treating, suppressing, reducing the severity, reducing the risk, or inhibiting
a drug resistant cancer
wherein the cancer is melanoma; e) a method of treating, suppressing, reducing
the severity,
reducing the risk, or inhibiting a drug resistant cancer wherein the cancer is
prostate cancer; f) a
method of treating, suppressing, reducing the severity, reducing the risk, or
inhibiting metastatic
92
CA 3030689 2019-01-18
melanoma; g) a method of treating, suppressing, reducing the severity,
reducing the risk, or
inhibiting prostate cancer; h) treating, suppressing, reducing the severity,
reducing the risk, or
inhibiting cancer in a subject, wherein the subject has been previously
treated with chemotherapy,
radiotherapy, or biological therapy; comprising the step of administering to
said subject a
compound of this invention and/or an isomer, metabolite, pharmaceutically
acceptable salt,
pharmaceutical product, tautomer, hydrate, N-oxide, polymorph, or crystal of
said compound, or
any combination thereof.
[00212] The compounds of the present invention are useful in the treatment,
reducing the severity,
reducing the risk, or inhibition of cancer, metastatic cancer, drug resistant
tumors, drug resistant
cancer and various forms of cancer. In a preferred embodiment the cancer is
prostate cancer, breast
cancer, ovarian cancer, skin cancer (e.g., melanoma), lung cancer, colon
cancer, leukemia,
lymphoma, head and neck, pancreatic, esophageal, renal cancer or CNS cancer
(e.g., glioma,
glioblastoma). Treatment of these different cancers is supported by the
Examples herein. Moreover,
based upon their believed mode of action as tubulin inhibitors, it is believed
that other forms of
cancer will likewise be treatable or preventable upon administration of the
compounds or
compositions of the present invention to a patient. Preferred compounds of the
present invention are
selectively disruptive to cancer cells, causing ablation of cancer cells but
preferably not normal cells.
Significantly, harm to normal cells is minimized because the cancer cells are
susceptible to
disruption at much lower concentrations of the compounds of the present
invention.
[00213] In some embodiments, this invention provides for the use of a compound
as herein
described, or its isomer, metabolite, pharmaceutically acceptable salt,
pharmaceutical product,
tautomer, polymorph, crystal, N-oxide, hydrate or any combination thereof, for
treating, suppressing,
reducing the severity, reducing the risk, or inhibiting cancer in a subject.
In another embodiment, the
" cancer is adrenocortical carcinoma, anal cancer, bladder cancer, brain
tumor, brain stem tumor,
breast cancer, glioma, cerebellar astrocytoma, cerebral astrocytoma,
ependymoma,
medulloblastoma, supratentorial primitive neuroectodermal, pineal tumors,
hypothalamic glioma,
breast cancer, carcinoid tumor, carcinoma, cervical cancer, colon cancer,
central nervous system
(CNS) cancer, endometrial cancer, esophageal cancer, extrahepatic bile duct
cancer, Ewing's family
of tumors (Pnet), extracranial germ cell tumor, eye cancer, intraocular
melanoma, gallbladder
cancer, gastric cancer, germ cell tumor, extragonadal, gestational
trophoblastic tumor, head and neck
cancer, hypopharyngeal cancer, islet cell carcinoma, laryngeal cancer,
leukemia, acute
93
CA 3030689 2019-01-18
lymphoblastic, leukemia, oral cavity cancer, liver cancer, lung cancer, non-
small cell lung cancer,
small cell, lymphoma, AIDS-related lymphoma, central nervous system (primary),
lymphoma,
cutaneous T-cell, lymphoma, Hodgkin's disease, non-Hodgkin's disease,
malignant mesothelioma,
melanoma, Merkel cell carcinoma, metasatic squamous carcinoma, multiple
myeloma, plasma cell
neoplasms, mycosis fungoides, myelodysplastic syndrome, myeloproliferative
disorders,
nasopharyngeal cancer, neuroblastoma, oropharyngeal cancer, osteosarcoma,
ovarian cancer,
ovarian epithelial cancer, ovarian germ cell tumor, ovarian low malignant
potential tumor, pancreatic
cancer, exocrine, pancreatic cancer, islet cell carcinoma, paranasal sinus and
nasal cavity cancer,
parathyroid cancer, penile cancer, pheochromocytoma cancer, pituitary cancer,
plasma cell
neoplasm, prostate cancer, rhabdomyosarcoma, rectal cancer, renal cancer,
renal cell cancer, salivary
gland cancer, Sezary syndrome, skin cancer, cutaneous T-cell lymphoma, skin
cancer, Kaposi's
sarcoma, skin cancer, melanoma, small intestine cancer, soft tissue sarcoma,
soft tissue sarcoma,
testicular cancer, thymoma, malignant, thyroid cancer, urethral cancer,
uterine cancer, sarcoma,
unusual cancer of childhood, vaginal cancer, vulvar cancer, Wilms' tumor, or
any combination
thereof. In another embodiment the subject has been previously treated with
chemotherapy,
radiotherapy or biological therapy.
[00214] In some embodiments, this invention provides for the use of a compound
as herein
described, or its isomer, metabolite, pharmaceutically acceptable salt,
pharmaceutical product,
tautomer, polymorph, crystal, N-oxide, hydrate or any combination thereof, for
treating, suppressing,
reducing the severity, reducing the risk, or inhibiting a metastatic cancer in
a subject. In another
embodiment, the cancer is adrenocortical carcinoma, anal cancer, bladder
cancer, brain tumor, brain
stem tumor, breast cancer, glioma, cerebellar astrocytoma, cerebral
astrocytoma, ependymoma, =
medulloblastoma, supratentorial primitive neuroectodermal, pineal tumors,
hypothalamic glioma,
breast cancer, carcinoid tumor, carcinoma, cervical cancer, colon cancer,
central nervous system
(CNS) cancer, endometrial cancer, esophageal cancer, extrahepatic bile duct
cancer, Ewing's family
of tumors (Pnet), extracranial germ cell tumor, eye cancer, intraocular
melanoma, gallbladder
cancer, gastric cancer, germ cell tumor, extragonadal, gestational
trophoblastic tumor, head and neck
cancer, hypopharyngeal cancer, islet cell carcinoma, laryngeal cancer,
leukemia, acute
lymphoblastic, leukemia, oral cavity cancer, liver cancer, lung cancer, non-
small cell lung cancer,
small cell, lymphoma, AIDS-related lymphoma, central nervous system (primary),
lymphoma,
cutaneous 1-cell, lymphoma, Hodgkin's disease, non-Hodgkin's disease,
malignant mesothelioma,
94
CA 3030689 2019-01-18
melanoma, Merkel cell carcinoma, metasatic squamous carcinoma, multiple
myeloma, plasma cell
neoplasms, mycosis fungoides, myelodysplastic syndrome, myeloproliferative
disorders,
nasopharyngeal cancer, neuroblastoma, oropharyngeal cancer, osteosarcoma,
ovarian cancer,
ovarian epithelial cancer, ovarian germ cell tumor, ovarian low malignant
potential tumor, pancreatic
cancer, exocrine, pancreatic cancer, islet cell carcinoma, paranasal sinus and
nasal cavity cancer,
parathyroid cancer, penile cancer, pheochromocytoma cancer, pituitary cancer,
plasma cell
neoplasm, prostate cancer, rhabdomyosarcoma, rectal cancer, renal cancer,
renal cell cancer, salivary
gland cancer, Sezary syndrome, skin cancer, cutaneous T-cell lymphoma, skin
cancer, Kaposi's
sarcoma, skin cancer, melanoma, small intestine cancer, soft tissue sarcoma,
soft tissue sarcoma,
testicular cancer, thymoma, malignant, thyroid cancer, urethral cancer,
uterine cancer, sarcoma,
unusual cancer of childhood, vaginal cancer, vulvar cancer, Wilms' tumor, or
any combination
thereof.
[00215] In some embodiments, this invention provides for the use of a compound
as herein
described, or its isomer, metabolite, pharmaceutically acceptable salt,
pharmaceutical product,
tautomer, polymorph, crystal, N-oxide, hydrate or any combination thereof, for
treating, suppressing,
reducing the severity, reducing the risk, or inhibiting a drug-resistant
cancer or resistant cancer in a
subject. In another embodiment, the cancer is adrenocortical carcinoma, anal
cancer, bladder cancer,
brain tumor, brain stem tumor, breast cancer, glioma, cerebellar astrocytoma,
cerebral astrocytoma,
ependyrnoma, medulloblastoma, supratentorial primitive neuroectodermal, pineal
tumors,
hypothalamic glioma, breast cancer, carcinoid tumor, carcinoma, cervical
cancer, colon cancer,
central nervous system (CNS) cancer, endometrial cancer, esophageal cancer,
extrahepatic bile duct
cancer, Ewing's family of tumors (Pnet), extracranial germ cell tumor, eye
canccr, intraoeular
melanoma, gallbladder cancer, gastric cancer, germ cell tumor, extragonadal,
gestational
trophoblastic tumor, head and neck cancer, hypopharyngeal cancer, islet cell
carcinoma, laryngeal
cancer, leukemia, acute lymphoblastic, leukemia, oral cavity cancer, liver
cancer, lung cancer, non-
small cell lung cancer, small cell, lymphoma, AIDS-related lymphoma, central
nervous system
(Primary), lymphoma, cutaneous T-cell, lymphoma, Hodgkin's disease, non-
Hodgkin's disease,
malignant mesothelioma, melanoma, Merkel cell carcinoma, metasatic squamous
carcinoma,
multiple myeloma, plasma cell neoplasms, mycosis fiingoides, myelodysplastic
syndrome,
myeloproliferative disorders, nasopharyngeal cancer, neuroblastoma,
oropharyngeal cancer,
osteosarcoma, ovarian cancer, ovarian epithelial cancer, ovarian germ cell
tumor, ovarian low
CA 3030689 2019-01-18
malignant potential tumor, pancreatic cancer, exocrine, pancreatic cancer,
islet cell carcinoma,
paranasal sinus and nasal cavity cancer, parathyroid cancer, penile cancer,
pheochromocytoma
cancer, pituitary cancer, plasma cell neoplasm, prostate cancer,
rhabdomyosarcoma, rectal cancer,
renal cancer, renal cell cancer, salivary gland cancer, Sezary syndrome, skin
cancer, cutaneous T-
cell lymphoma, skin cancer, Kaposi's sarcoma, skin cancer, melanoma, small
intestine cancer, soft
tissue sarcoma, soft tissue sarcoma, testicular cancer, thymoma, malignant,
thyroid cancer, urethral
cancer, uterine cancer, sarcoma, unusual cancer of childhood, vaginal cancer,
vulvar cancer, Wilms'
tumor, or any combination thereof.
[00216] In one embodiment "metastatic cancer" refers to a cancer that spread
(metastasized) from
its original site to another area of the body. Virtually all cancers have the
potential to spread.
Whether metastases develop depends on the complex interaction of many tumor
cell factors,
including the type of cancer, the degree of maturity (differentiation) of the
tumor cells, the location
and how long the cancer has been present, as well as other incompletely
understood factors.
Metastases spread in three ways - by local extension from the tumor to the
surrounding tissues,
through the bloodstream to distant sites or through the lymphatic system to
neighboring or distant
lymph nodes. Each kind of cancer may have a typical route of spread. The tumor
is called by the
primary site (ex. breast cancer that has spread to the brain is called
metastatic breast cancer to the
brain).
[00217] In one embodiment "drug-resistant cancer" refers to cancer
cells that acquire
resistance to chemotherapy. Cancer cells can acquire resistance to
chemotherapy by a range of
mechanisms, including the mutation or overexpression of the drug target,
inactivation of the drug,
or elimination of the drug from the cell. Tumors that recur after an initial
response to
chemotherapy may be resistant to multiple drugs (they are multidrug
resistant). In the
conventional view of drug resistance, one or several cells in the tumor
population acquire genetic
changes that confer drug resistance. Accordingly, the reasons for drug
resistance, inter alia, are:
a) some of the cells that are not killed by the chemotherapy mutate (change)
and become resistant
to the drug. Once they multiply, there may be more resistant cells than cells
that are sensitive to
the chemotherapy; b) Gene amplification. A cancer cell may produce hundreds of
copies of a
particular gene. This gene triggers an overproduction of protein that renders
the anticancer drug
ineffective; c) cancer cells may pump the drug out of the cell as fast as it
is going in using a
molecule called p-glycoprotein; d) cancer cells may stop taking in the drugs
because the protein
96
CA 3030689 2019-01-18
that transports the drug across the cell wall stops working; e) the cancer
cells may learn how to
repair the DNA breaks caused by some anti-cancer drugs; f) cancer cells may
develop a
mechanism that inactivates the drug. One major contributor to multidrug
resistance is
overexpression of P-glycoprotein (P-gp). This protein is a clinically
important transporter protein
belonging to the ATP-binding cassette family of cell membrane transporters. It
can pump
substrates including anticancer drugs out of tumor cells through an ATP-
dependent mechanism.
Thus, the resistance to anticancer agents used in chemotherapy is the main
cause of treatment
failure in malignant disorders, provoking tumors to become resistant. Drug
resistance is the major
cause of cancer chemotherapy failure.
100218] In one embodiment "resistant cancer" refers to drug-resistant
cancer as described
herein above. In another embodiment "resistant cancer" refers to cancer cells
that acquire
resistance to any treatment such as chemotherapy, radiotherapy or biological
therapy.
[00219] In one embodiment, this invention is directed to treating,
suppressing, reducing the
severity, reducing the risk, or inhibiting cancer in a subject, wherein the
subject has been
previously treated with chemotherapy, radiotherapy or biological therapy.
[00220] In one embodiment "Chemotherapy" refers to chemical treatment
for cancer such
as drugs that kill cancer cells directly. Such drugs are referred as "anti-
cancer" drugs or
"antineoplastics." Today's therapy uses more than 100 drugs to treat cancer.
To cure a specific
cancer. Chemotherapy is used to control tumor growth when cure is not
possible; to shrink
tumors before surgery or radiation therapy; to relieve symptoms (such as
pain); and to destroy
microscopic cancer cells that may be present after the known tumor is removed
by surgery (called
adjuvant therapy). Adjuvant therapy is given to prevent a possible cancer
reoccurrence.
[002211 In one embodiment, "Radiotherapy" refers to high energy x-rays
and similar rays
(such as electrons) to treat disease. Many people with cancer will have
radiotherapy as part of
their treatment. This can be given either as external radiotherapy from
outside the body using x-
rays or from within the body as internal radiotherapy. Radiotherapy works by
destroying the
cancer cells in the treated area. Although normal cells can also be damaged by
the radiotherapy,
they can usually repair themselves. Radiotherapy treatment can cure some
cancers and can also
reduce the chance of a cancer coming back after surgery. It may be used to
reduce cancer
symptoms.
97
CA 3030689 2019-01-18
[00222] In one embodiment "Biological therapy" refers to substances
that occur naturally
in the body to destroy cancer cells. There are several types of treatment
including: monoclonal
antibodies, cancer growth inhibitors, vaccines and gene therapy. Biological
therapy is also known
as immunotherapy.
[00223] In one embodiment, this invention provides a method of treating a
subject
suffering from prostate cancer, metastatic prostate cancer, resistant prostate
cancer or drug-
resistant prostate cancer comprising the step of administering to said subject
a compound of this
invention, or its isomer, metabolite, pharmaceutically acceptable salt,
pharmaceutical product,
tautomer, hydrate, N-oxide, polymorph, crystal or any combination thereof, or
a composition
comprising the same in an amount effective to treat prostate cancer in the
subject. In another
embodiment, the compound is compound 12db. In another embodiment, the compound
is
compound llcb. In another embodiment, the compound is compound lift. In
another
embodiment, the compound is compound 12da. In another embodiment, the compound
is
compound 12fa. In another embodiment, the compound is compound 12fb. In
another
embodiment, the compound is compound 12cb. In another embodiment, the compound
is
compound 55. In another embodiment, the compound is compound 6b. In another
embodiment,
the compound is compound 17ya.
[00224] In one embodiment, this invention provides a method for
suppressing, reducing
the severity, reducing the risk, delaying the progression, or inhibiting
prostate cancer, metastatic
prostate cancer, resistant prostate cancer or drug-resistant prostate cancer
in a subject, comprising
administering to the subject a compound of this invention and/or its isomer,
metabolite,
pharmaceutically acceptable salt, pharmaceutical product, tautomer, hydrate, N-
oxide,
polymorph, crystal or any combination thereof or a composition comprising the
same. In another
embodiment, the compound is compound 12db. In another embodiment, the compound
is
compound llcb. In another embodiment, the compound is compound lift. In
another
embodiment, the compound is compound 12da. In another embodiment, the compound
is
compound 12fa. In another embodiment, the compound is compound 12fb. In
another
embodiment, the compound is compound 12cb. In another embodiment, the compound
is
compound SS. In another embodiment, the compound is compound 6b. In another
embodiment,
the compound is compound 17ya.
98
CA 3030689 2019-01-18
[00225] In one embodiment, this invention provides a method of
treating a subject
suffering from breast cancer, metastatic breast cancer, resistant breast
cancer or drug-resistant
breast cancer comprising the step of administering to said subject a compound
of this invention,
or its isomer, metabolite, pharmaceutically acceptable salt, pharmaceutical
product, tautomer,
hydrate, N-oxide, polymorph, crystal or any combination thereof, or a
composition comprising
the same. In another embodiment, the subject is a male or female. In another
embodiment, the
compound is compound 12db. In another embodiment, the compound is compound
11cb. In
another embodiment, the compound is compound 11113. In another embodiment, the
compound is
compound 12da. In another embodiment, the compound is compound 12fa. In
another
embodiment, the compound is compound 12fb. In another embodiment, the compound
is
compound 12cb. In another embodiment, the compound is compound 55. In another
embodiment, the compound is compound 6b. In another embodiment, the compound
is
compound 17ya.
[00226] In one embodiment, this invention provides a method of
suppressing, reducing the
severity, reducing the risk, delaying the progression, or inhibiting breast
cancer, metastatic breast
cancer, resistant breast cancer or drug-resistant breast cancer in a subject
comprising the step of
administering to said subject a compound of this invention or its isomer,
metabolite,
pharmaceutically acceptable salt, pharmaceutical product, tautomer,hydrate, N-
oxide, polymorph,
crystal or any combination thereof, or a composition comprising the same. In
another
embodiment, the subject is a male or female. In another embodiment, the
compound is
compound 12db. In another embodiment, the compound is compound llcb. In
another
embodiment, the compound is compound 111M. In another embodiment, the compound
is
compound 12da. In another embodiment, the compound is compound 12fa. In
another
embodiment, the compound is compound 12th. In another embodiment, the compound
is
compound 12cb. In another embodiment, the compound is compound 55. In another
embodiment, the compound is compound 6h. In another embodiment, the compound
is
compound 17ya.
[002271 In another embodiment, this invention provides for the use of
a compound as
herein described, or its isomer, metabolite, pharmaceutically acceptable salt,
pharmaceutical
- 30 product, tautomer, hydrate, N-oxide, polymorph, crystal or any
combination thereof, for treating,
suppressing, reducing the severity, reducing the risk, delaying the
progression, or inhibiting
99
CA 3030689 2019-01-18
ovarian cancer, metastatic ovarian cancer, resistant ovarian cancer or drug-
resistant ovarian
cancer in a subject. In another embodiment, the compound is compound 12db. In
another
embodiment, the compound is compound 11cb. In another embodiment, the compound
is
compound llfb. In another embodiment, the compound is compound 12da. In
another
embodiment, the compound is compound 12fa. In another embodiment, the compound
is
compound 12fb. In another embodiment, the compound is compound 12th. In
another
embodiment, the compound is compound 55. In another embodiment, the compound
is
compound 6b. In another embodiment, the compound is compound 17ya.
[00228] In one embodiment, this invention provides a method for
treating, suppressing,
reducing the severity, reducing the risk or inhibiting melanoma, metastatic
melanoma, resistant
melanoma or drug-resistant melanoma in a subject, comprising administering to
the subject a
- compound of this invention and/or its isomer, metabolite, pharmaceutically
acceptable salt,
pharmaceutical product, tautomer, hydrate, N-oxide, polymorph, crystal or any
combination
thereof. In another embodiment, the compound is compound 12db. In another
embodiment, the
compound is compound 11th. In another embodiment, the compound is compound
11th. In
another embodiment, the compound is compound 12da. In another embodiment, the
compound is
compound 12fa. In another embodiment, the compound is compound 12th. In
another
embodiment, the compound is compound 12eb. In another embodiment, the compound
is
compound 55. In another embodiment, the compound is compound 6b. In another
embodiment,
the compound is compound 17ya.
[00229] In another embodiment, this invention provides for the use of a
compound as
herein described, or isomer, metabolite, pharmaceutically acceptable salt,
pharmaceutical
product, tautomer, hydrate, N-oxide, polymorph, crystal or any combination
thereof, for treating,
suppressing, reducing the severity, reducing the risk, delaying the
progression, or inhibiting lung
cancer, metastatic lung cancer, resistant lung cancer or drug-resistant lung
cancer. In another
embodiment, the compound is compound 12db. In another embodiment, the compound
is
compound llcb. In another embodiment, the compound is compound 11th. In
another
embodiment, the compound is compound 12da. In another embodiment, the compound
is
compound 12fa. In another embodiment, the compound is compound 12th. In
another
embodiment, the compound is compound 12cb. In another embodiment, the compound
is
100
CA 3030689 2019-01-18
compound 55. In another embodiment, the compound is compound 6b. In another
embodiment,
the compound is compound 17ya.
[00230] In another embodiment, this invention provides for the use of a
compound as
herein described, or isomer, metabolite, pharmaceutically acceptable salt,
pharmaceutical
product, tautomer, hydrate, N-oxide, polymorph, crystal any combination
thereof, for treating,
suppressing, reducing the severity, reducing the risk, delaying the
progression, or inhibiting non-
small cell lung cancer, metastatic small cell lung cancer, resistant small
cell lung cancer or drug-
resistant small cell lung cancer. In another embodiment, the compound is
compound 12db. In
another embodiment, the compound is compound llcb. In another embodiment, the
compound is
compound llfb. In another embodiment, the compound is compound 12da. In
another
embodiment, the compound is compound 12fa. In another embodiment, the compound
is
compound 12fb. In another embodiment, the compound is compound 12cb. In
another
embodiment, the compound is compound 55. In another embodiment, the compound
is
compound 6b. In another embodiment, the compound is compound 17ya.
[00231] In another embodiment, this invention provides for the use of a
compound as
herein described, or isomer, metabolite, pharmaceutically acceptable salt,
pharmaceutical
product, tautomer, hydrate, N-oxide, polymorph, crystal any combination
thereof, for treating,
suppressing, reducing the severity, reducing the risk, delaying the
progression, or inhibiting colon
cancer, metastatic colon cancer, resistant colon cancer or drug-resistant
colon cancer. In another
embodiment, the compound is compound 12db. In another embodiment, the compound
is
compound Ilcb. In another embodiment, the compound is compound 11th. In
another
embodiment, the compound is compound 12cla. In another embodiment, the
compound is
compound 12fa. In another embodiment, the compound is compound 12113. In
another
embodiment, the compound is compound 12cb. In another embodiment, the compound
is
compound 55. In another embodiment, the compound is compound 6b. In another
embodiment,
the compound is compound 17ya.
[00232] In another embodiment, this invention provides for the use of a
compound as
herein described, or isomer, metabolite, pharmaceutically acceptable salt,
pharmaceutical
product, tautomer, hydrate, N-oxide, polymorph, crystal any combination
thereof, for treating,
suppressing, reducing the severity, reducing the risk, delaying the
progression, or inhibiting of
leukemia, metastatic leukemia, resistant leukemia or drug-resistant leukemia.
In another
101
CA 3030689 2019-01-18
embodiment, the compound is compound 12db. In another embodiment, the compound
is
compound llcb. In another embodiment, the compound is compound llfb. In
another
embodiment, the compound is compound 12da. In another embodiment, the compound
is
compound 12fa. In another embodiment, the compound is compound 12Th. In
another
embodiment, the compound is compound 12cb. In another embodiment, the compound
is
compound 55. In another embodiment, the compound is compound 6b. In another
embodiment,
the compound is compound 17ya.
[00233] In
another embodiment, this invention provides for the use of a compound as
herein described, or isomer, metabolite, pharmaceutically acceptable salt,
pharmaceutical
product, tautomer,. hydrate, N-oxide, polymorph, crystal any combination
thereof, for treating,
suppressing, reducing the severity, reducing the risk, delaying the
progression, or inhibiting
lymphoma, metastatic lymphoma, lymphoma or drug-resistant lymphoma. In
another
embodiment, the compound is compound 12db. In another embodiment, the compound
is
compound llcb. In another embodiment, the compound is compound 11Th. In
another
embodiment, the compound is compound 12da. In another embodiment, the compound
is
compound 12fa. In another embodiment, the compound is compound 12Th. In
another
embodiment, the compound is compound 12cb. In another embodiment, the compound
is
compound 55. In another embodiment, the compound is compound 6b. In another
embodiment,
the compound is compound 17ya.
[00234] In another
embodiment, this invention provides for the use of a compound as
herein described, or isomer, metabolite, pharmaceutically acceptable salt,
pharmaceutical
product, tautomer, hydrate, N-oxide, polymorph, crystal any combination
thereof, for treating,
suppressing, reducing the severity, reducing the risk, delaying the
progression, or inhibiting head
and neck cancer, metastatic head and neck cancer, resistant head and neck
cancer or drug-
resistant head and neck cancer. In another embodiment, the compound is
compound 12db. In
another embodiment, the compound is compound llcb. In another embodiment, the
compound is
compound llfb. In another embodiment, the compound is compound 12da. In
another
embodiment, the compound is compound 12fa. In another embodiment, the compound
is
compound 12Th. In another embodiment, the compound is compound 12th. In
another
embodiment, the compound is compound 55. In another embodiment, the compound
is
compound 6b. In another embodiment, the compound is compound 17ya.
102
=
CA 3030689 2019-01-18
[00235] In another embodiment, this invention provides for the use of a
compound as
herein described, or isomer, metabolite, pharmaceutically acceptable salt,
pharmaceutical
product, tautomer, hydrate, N-oxide, polymorph, crystal any combination
thereof, for treating,
suppressing, reducing the severity, reducing the risk, delaying the
progression, or inhibiting of
pancreatic cancer, metastatic pancreatic cancer, resistant pancreatic cancer
or drug-resistant
pancreatic cancer. In another embodiment, the compound is compound 12db. In
another
embodiment, the compound is compound llcb. In another embodiment, the compound
is
compound 11th. In another embodiment, the conipound is compound 12da. In
another
embodiment, the compound is compound 12fa. In another embodiment, the compound
is
compound 12M. In another embodiment, the compound is compound 12cb. In another
embodiment, the compound is compound 55. In another embodiment, the compound
is
compound 6b. In another embodiment, the compound is compound 17ya.
[002361 In another embodiment, this invention provides for the use of a
compound as
herein described, or isomer, metabolite, pharmaceutically acceptable salt,
pharmaceutical
product, tautomer, hydrate, N-oxide, polymorph, crystal any combination
thereof, for treating,
suppressing, reducing the severity, reducing the risk, delaying the
progression, or inhibiting
esophageal cancer, metastatic esophageal cancer, resistant esophageal cancer
or drug-resistant
esophageal cancer. In another embodiment, the compound is compound 12db. In
another
embodiment, the compound is compound llcb. In another embodiment, the compound
is
compound llfb. In another embodiment, the compound is compound 12da. In
another -
embodiment, the compound is compound 12fa. In another embodiment, the compound
is
compound 12th. In another embodiment, the compound is compound 120. In another
embodiment, the compound is compound 55. In another embodiment, the compound
is
compound 6b. In another embodiment, the compound is compound 17ya.
[00237] In another embodiment, this invention provides for the use of a
compound as
herein described, or isomer, metabolite, pharmaceutically acceptable salt,
pharmaceutical
product, tautomer, hydrate, N-oxide, polymorph, crystal any combination
thereof, for treating,
suppressing, reducing the severity, reducing the risk, delaying the
progression, or inhibiting renal
cancer, Metastatic renal cancer, resistant renal cancer or drug-resistant
renal cancer. In another
embodiment, the compound is compound 12db. In another embodiment, the compound
is
compound llcb. In another embodiment, the compound is compound 11Th. In
another
103
CA 3030689 2019-01-18
embodiment, the compound is compound 12da. In another embodiment, the compound
is
compound 12fa. In another embodiment, the compound is compound 12fb. In
another
embodiment, the compound is compound 12cb. In another embodiment, the compound
is
compound 55. In another embodiment, the compound is compound 6b. In another
embodiment,
the compound is compound 17ya.
[00238] In another embodiment, this invention provides for the use of a
compound as
herein described, or isomer, metabolite, pharmaceutically acceptable salt,
pharmaceutical
product, tautomer, hydrate, N-oxide, polymorph, crystal any combination
thereof, for treating,
suppressing, reducing the severity, reducing the risk, delaying the
progression, or inhibiting CNS
cancer, metastatic CNS cancer, resistant CNS cancer or drug-resistant CNS
cancer. In another
embodiment, the compound is compound 12db. In another embodiment, the compound
is
compound llcb. In another embodiment, the compound is compound 1103. In
another
embodiment, the compound is compound 12da. In another embodiment, the compound
is
compound 12fa. In another embodiment, the compound is compound 12Th. In
another
embodiment, the compound is compound 12cb. In another embodiment, the compound
is
compound 55. In another embodiment, the compound is compound 6b. In another
embodiment,
the compound is compound 17ya.
[00239] In some embodiments, this invention provides for the use of a compound
as herein
described, or its isomer, metabolite, pharmaceutically acceptable salt,
pharmaceutical product,
tautomer, polymorph, crystal, N-oxide, hydrate or any combination thereof, for
treating, suppressing,
reducing the severity, reducing the risk, or inhibiting a drug resistant
cancerous tumor or tumors in a
subject. In another embodiment, the cancer is adrenocortical carcinoma, anal
cancer, bladder cancer,
brain tumor, brain stem tumor, breast cancer, glioma, cerebellar astrocytoma,
cerebral astrocytoma,
ependymoma, medulloblastoma, supratentorial primitive neuroectodermal, pineal
tumors,
hypothalamic glioma, breast cancer, carcinoid tumor, carcinoma, cervical
cancer, colon cancer,
central nervous system (CNS) cancer, endometrial cancer, esophageal cancer,
extrahepatic bile duct
cancer, Ewing's family of tumors (Pnet), extracranial germ cell tumor, eye
cancer, intraocular
melanoma, gallbladder cancer, gastric cancer, germ cell tumor, extragonadal,
gestational
trophoblastic tumor, head and neck cancer, hypopharyngeal cancer, islet cell
carcinoma, laryngeal
cancer, leukemia, acute lymphoblastic, leukemia, oral cavity cancer, liver
cancer, lung cancer, non-
small cell lung cancer, small cell, lymphoma, AIDS-related lymphoma, central
nervous system
104
CA 3030689 2019-01-18
(primary), lymphoma, cutaneous T-cell, lymphoma, Hodgkin's disease, non-
Hodgkin's disease,
malignant mesothelioma, melanoma, Merkel cell carcinoma, metasatic squamous
carcinoma,
multiple myeloma, plasma cell neoplasms, mycosis fungoides, myelodysplastic
syndrome,
myeloproliferative disorders, nasopharyngeal cancer, neuroblastoma,
oropharyngeal cancer,
osteosarcoma, ovarian cancer, ovarian epithelial cancer, ovarian germ cell
tumor, ovarian low
malignant potential tumor, pancreatic cancer, exocrine, pancreatic cancer,
islet cell carcinoma,
paranasal sinus and nasal cavity cancer, parathyroid cancer, penile cancer,
pheochromocytoma
cancer, pituitary cancer, plasma cell neoplasm, prostate cancer,
rhabdomyosarcoma, rectal cancer,
renal cancer, renal cell cancer, salivary gland cancer, Sezary syndrome, skin
cancer, cutaneous 1-
cell lymphoma, skin cancer, Kaposi's sarcoma, skin cancer, melanoma, small
intestine cancer, soft
tissue sarcoma, soft tissue sarcoma, testicular cancer, thyrnoma, malignant,
thyroid cancer, urethral
cancer, uterine cancer, sarcoma, unusual cancer of childhood, vaginal cancer,
vulvar cancer, Wilms'
tumor, or any combination thereof. In another embodiment, the compound is
compound 12db. In
another embodiment, the compound is compound 110. In another embodiment, the
compound is
compound 11f1). In another embodiment, the compound is compound 12da_ In
another
embodiment, the compound is compound 12fa. In another embodiment, the compound
is compound
12Th. In another embodiment, the compound is compound 12th. In another
embodiment, the
compound is compound 55. In another embodiment, the compound is compound 6b.
In another
embodiment, the compound is compound 17ya.
[00240] In
another embodiment, the tumor is prostate cancer tumor. In another
embodiment, the tumor is ovarian cancer tumor. In another embodiment, the
tumor is a
melanoma tumor. In another embodiment, the tumor is a multidrug resistant
(MDR) melanoma
tumor.
[00241] In one embodiment, this invention is directed to a method of
destroying a cancerous cell
comprising: providing a compound of this invention and contacting the
cancerous cell with the
compound under conditions effective to destroy the contacted cancerous cell.
According to various
embodiments of destroying the cancerous cells, the cells to be destroyed can
be located either in vivo
or ex vivo (i.e., in culture). In another embodiment, the compound is compound
12db. In another
embodiment, the compound is compound 110. In another embodiment, the compound
is compound
11Th. In another embodiment, the compound is compound 12da. In another
embodiment, the
compound is compound 12fa. In another embodiment, the compound is compound
1211). In another
105
CA 3030689 2019-01-18
embodiment, the compound is compound 12eb. In another embodiment, the compound
is compound
55. In another embodiment, the compound is compound 6b. In another embodiment,
the compound
is compound 17ya.
[00242] In another embodiment, the cancer is selected from the group
consisting of prostate
cancer, breast cancer, ovarian cancer, skin cancer, melanoma, lung cancer,
colon cancer, leukemia,
renal cancer, CNS cancer, and combinations thereof.
[00243] A
still further aspect of the present invention relates to a method of treating
or
preventing a cancerous condition that includes: providing a compound of the
present invention
and then administering an effective amount of the compound to a patient in a
manner effective to.
.. treat or prevent a cancerous condition.
[00244]
According to one embodiment, the patient to be treated is characterized by the
presence of a precancerous condition, and the administering of the compound is
effective to
prevent development of the precancerous condition into the cancerous
condition. This can occur
by destroying the precancerous cell prior to or concurrent with its further
development into a
cancerous state.
[00245]
According to another embodiment, the patient to be treated is characterized by
the presence of a cancerous condition, and the administering of the compound
is effective either
to cause regression of the cancerous condition or to inhibit growth of the
cancerous condition,
i.e., stopping its growth altogether or reducing its rate of growth. This
preferably occurs by
destroying cancer cells, regardless of their location in the patient body.
That is, whether the
cancer cells are located at a primary tumor site or whether the cancer cells
have metastasized and
created secondary tumors within the patient body.
[00246] As
used herein, subject or patient refers to any mammalian patient, including
without limitation, humans and other primates, dogs, cats, horses, cows,
sheep, pigs, rats, mice,
and other rodents. In one embodiment, the subject is male. In another
embodiment, the subject is
female. In some embodiments, while the methods as described herein may be
useful for treating
either males or females.
[00247] When
administering the compounds of the present invention, they can be
administered systemically or, alternatively, they can be administered directly
to a specific site
where cancer cells or precancerous cells are present. Thus, administering can
be accomplished in
106
CA 3030689 2019-01-18
any manner effective for delivering the compounds or the pharmaceutical
compositions to the
cancer cells or precancerous cells. Exemplary modes of administration include,
without .
limitation, administering the compounds or compositions orally, topically,
transdermally,
parenterally, subcutaneously, intravenously, intramuscularly,
intraperitoneally, by intranasal
instillation, by intracavitary or intravesical instillation, intraocularly,
intraarterially,
intralesionally, or by application to mucous membranes, such as, that of the
nose, throat, and
bronchial tubes.
[00248) The compounds of the present invention are useful in the
treatment or prevention
of various forms of cancer, particularly prostate cancer, breast cancer,
ovarian, skin cancer (e.g.,
melanoma), lung cancer, colon cancer, leukemia, renal cancer, and CNS cancer
(e.g., glioma,
glioblastoma). Treatment of these different cancers is supported by the
Examples herein.
Moreover, based upon their believed mode of action as tubulin inhibitors, it
is believed that other
forms of cancer will likewise be treatable or preventable upon administration
of the compounds
or compositions of the present invention to a patient. Preferred compounds of
the present
invention are selectively disruptive to cancer cells, causing ablation of
cancer cells but preferably
not normal cells. Significantly, harm to normal cells is minimized because the
cancer cells are
susceptible to disruption at much lower concentrations of the compounds of the
present
invention.
[00249] The compounds of the present invention are useful in the treatment,
reducing the severity,
reducing the risk, or inhibition of cancer, metastatic cancer, resistant
cancer or drug-resistant cancer.
In another embodiment, the cancer is prostate cancer, breast cancer, ovarian,
skin cancer (e.g.,
melanoma), lung cancer, colon cancer, leukemia, lymphoma, head and neck,
pancreatic, esophageal,
renal cancer or CNS cancer. Treatment of these different cancers is supported
by the Examples
herein. Moreover, based upon their believed mode of action as tubulin
inhibitors, it is believed that
other forms of cancer will likewise be treatable or preventable upon
administration of the
compounds or compositions of the present invention to a patient. Preferred
compounds of the
present invention are selectively disruptive to cancer cells, causing ablation
of cancer cells but
preferably not normal cells. Significantly, harm to normal cells is minimized
because the cancer
cells are susceptible to disruption at much lower concentrations of the
compounds of the present
invention. In another embodiment, the compound is compound 12db. In another
embodiment, the
compound is compound licb. In another embodiment, the compound is compound
llfb. In another
107
CA 3030689 2019-01-18
embodiment, the compound is compound 12da. In another embodiment, the compound
is compound
121a. In another embodiment, the compound is compound 12M. In another
embodiment, the =
compound is compound 12cb. In another embodiment, the compound is compound 55.
In another
embodiment, the compound is compound 6b. In another embodiment, the compound
is compound
17ya.
[00250] As used herein, subject or patient refers to any mammalian
patient, including
without limitation, humans and other primates, dogs, cats, horses, cows,
sheep, pigs, rats, mice,
and other rodents. In some embodiments, while the methods as described herein
may be useful
for treating either males or females.
[00251] In one embodiment, the compound is administered in combination with
an anti-
cancer agent by administering the compounds as herein described, alone or in
combination with
other agents.
[00252] When the compounds or pharmaceutical compositions of the
present invention are
administered to treat, suppress, reduce the severity, reduce the risk, or
inhibit a cancerous
condition, the pharmaceutical composition can also contain, or can be
administered in
conjunction with, other therapeutic agents or treatment regimen presently
known or hereafter
developed for the treatment of various types of cancer. Examples of other
therapeutic agents or
treatment regimen include, without limitation, radiation therapy,
immunotherapy, chemotherapy,
surgical intervention, and combinations thereof.
[00253] The following examples are presented in order to more fully illustrate
the preferred
embodiments of the invention. They should in no way, however, be construed as
limiting the broad
scope of the invention.
EXAMPLES
[00254] The Examples set forth below are for illustrative purposes only and
are not
intended to limit, in any way, the scope of the present invention.
Materials and Methods:
108
CA 3030689 2019-01-18
[00255] General. All reagents were purchased from Sigma-Aldrich
Chemical Co., Fisher
Scientific (Pittsburgh, PA), AK Scientific (Mountain View, CA), Oakwood
Products (West
Columbia, SC), etc. and were used without further purification. Moisture-
sensitive reactions were
carried under an argon atmosphere. ABT-751 was prepared according methods
reported by
Yoshino et at.26 Routine thin layer chromatography (TLC) was performed on
aluminum backed
Uniplates (Analtech, Newark, DE). Melting points were measured with Fisher-
Johns melting
point apparatus (uncorrected). NMR spectra were obtained on a Bruker AX 300
(Billerica, MA)
spectrometer or Varian Inova-500 (Vernon Hills, Illinois) spectrometer.
Chemical shifts are
reported as parts per million (ppm) relative to T1V1S in CDC13. Mass spectral
data was collected
on a Broker ESQUIRE electrospray/ion trap instrument in positive and negative
ion modes.
Elemental analyses were performed by Atlantic Microlab Inc.
[00256] Cell Culture and Cytotoxicity Assay of Prostate Cancer and
Melanoma. All
cell lines were obtained from ATCC (American Type Culture Collection,
Manassas, VA, USA),
while cell culture supplies were purchased from Cellgro Mediatech (Herndon,
VA, USA). We
examined the antiproliferative activity of our anti-tubulin compounds in four
human prostate
cancer cell lines (LNCaP, DU 145, PC-3, and PPC-1) and two human melanoma cell
lines (A375
and WM-164). Human ovarian cell line OVCAR-8 and its resistant cell line that
over-expresses
P-gp (NCl/ADR-RES) were used as MDR models. Both ovarian cell lines were
obtained from
National Cancer Institutes (NCI). All cell lines were tested and authenticated
by either ATCC or
NCI. All prostate cancer and ovarian cancer cell lines were cultured in RPMI
1640, supplemented
with 10% fetal bovine serum (FBS). Melanoma cells were cultured in DMEM,
supplemented
with 5% FBS, 1% antibiotic/antimycotic mixture (Sigma-Aldrich, Inc., Si Louis,
MO, USA) and
bovine insulin (5 i.tg/mL; Sigma-Aldrich). The cytotoxic potential of the anti-
tubulin compounds
was evaluated using the sulforhodamine B (SRB) assay after 96 h of treatment.
[00257] Aqueous Solubility. The solubility of drugs was determined by
Multiscreen
Solubility Filter Plate (Millipore Corporate, Billerica, MA) coupled with LC-
MS/MS. Briefly,
198 pl of phosphate buffered saline (PBS) buffer (pH 7.4) was loaded into 96-
well plate, and 2
.1.. of 10 mM test compounds (in DMSO) was dispensed and mixed with gentle
shaking (200-
300 rpm) for 1.5 h at RT (N = 3). The plate was centrifuged at 800g for 5 min,
and the filtrate
was used to determine its concentration and solubility of test compound by LC-
MS/MS as
described below.
109
CA 3030689 2019-01-18
[00258] Pharmacokinetic Study. Female Sprague-Dawley rats (n = 3 or 4; 254 4
g) were
purchased from Harlan Inc. (Indianapolis, IN). Rat thoracic jugular vein
catheters were
purchased from Braintree Scientific Inc. (Braintree, MA). On arrival at the
animal facility, the
animals were acclimated for 3 days in a temperature-controlled room (20-22 C)
with a 12 h
light/dark cycle before any treatment. Compound lh was administered
intravenously (i.v.) into
the jugular vein catheters at a dose of 2.5 mg/kg (in DMSO/PEG300, 2/8),
whereas 5Ha and 5Ik
were dosed at 5 mg/kg (in DMSO/PEG300, 1/9). An equal volume of heparinized
saline was
injected to replace the removed blood, and blood samples (250 pt) were
collected via the jugular
vein catheters at 10, 20, 30 min, and 1, 2, 4, 8, 12, 24 h. Compounds lh, 5Ha
and 5Hc were
given (p.o.) by oral gavage at 10 mg/kg (in Tweeem80/DMSO/1120, 2/1/7). All
blood samples
(250 lit) after oral administration were collected via the jugular vein
catheters at 30, 60, 90 mm,
120 min, 150 min, 180 min, 210 min, 240 min, and 8, 12, 24 h. Heparinized
syringes and vials
were prepared prior to blood collection. Plasma samples were prepared by
centrifuging the blood
samples at 8,000 g for 5 min. All plasma samples were stored immediately at -
80 C until
analyzed.
[00259] Analytes were extracted from 100 pt of plasma with 200 1.tL of
acetonitrile containing
200 nM the internal standard ((3,5-dimethoxyphenyl)(2-pheny1-1H-imidazol-4-
yOmethanone).
The samples were thoroughly mixed, centrifuged, and the organic extract was
transferred to
autosampler for LC-MS/MS analysis. Multiple reaction monitoring (MRM) mode,
scanning m/z
356 188 (compound 1h), m/z 371 203 (compound 5Ha), m/z 389 221 (compound 5Hc),
and m/z 309 171 (the internal standard), was used to obtain the most
sensitive signals. The
pharmaeokinetic parameters were determined using non-compartmental analysis
(WinNonlin,
Pharsight Corporation, Mountain View, CA).
[00260] Analytical Method. Sample solution (10 uL) was injected into an
Agilent series HPLC
system (Agilent 1100 Series Agilent 1100 Chernstation, Agilent Technology Co,
Ltd). All
analytes were separated on a narrow-bore C18 column (Alltech Alltima HP,
2.1x100 mm, 3 um,
Fisher, Fair Lawn, NJ). Two gradient modes were used. Gradient mode was used
to achieve the
separation of analytes using mixtures of mobile phase A [ACN/H20 (5%/95%, v/v)
containing
110
CA 3030689 2019-01-18
0.1% formic acid] and mobile phase B [ACW1-120 (95%/5%, v/v) containing 0.1%
formic acid]
at a flow rate of 300 tL/min. Mobile phase A was used at 15% from 0 to 1 min
followed by a
linearly programmed gradient to 100% of mobile phase B within 6 min, 100% of
mobile phase B
110a
CA 3030689 2019-01-18
was maintained for 0.5 min before a quick ramp to 15% mobile phase A. Mobile
phase A was
continued for another 12 mM towards the end of analysis.
[00261] In Vitro Tubulin Polymerization Assay. Bovine brain tubulin
(0.4 mg, >97%
pure) (Cytoskeleton, Denver, CO) was mixed with 10 M of the test compounds and
incubated in
100 gLof general tubulin buffer (80 mM PIPES, 2.0 mM MgC12, 0.5 mM EGTA, and 1
mM
GTP) at pH 6.9. The absorbance of wavelength at 340 nm was monitored every 1
min for 20 min
by the SYNERGY 4 Microplate Reader (Bio-Tek Instruments, Winooski, VT). The
spectrophotometer was set at 37 C for tubulin polymerization.
[00262] A triple-quadruple mass spectrometer, API Qtrap .40001-m
(Applied
Biosystems/MDS SCEEX, Concord, Ontario, Canada), operating with a
TurboIonSpray source
was used. The spraying needle voltage was set at 5 kV for positive mode.
Curtain gas was set at
10; Gas 1 and gas 2 were set 50. Collision-Assisted-Dissociation (CAD) gas at
medium and the
source heater probe temperature at 500 C. Data acquisition and quantitative
processing were
accomplished using AnalystTm software, Vet. 1.4.1 (Applied Biosystems).
The purity of the final compounds was tested via RP-HPLC on a Waters 2695 HPLC
system installed with a Photodiode Array Detector. Two RP-HPLC methods were
conducted
using a Supelco AscentisTm5pM C-18 column (250 x 4.6 mm) at ambient
temperature, and a
flow rate of 0.7 mUmin. HPLC1: Gradient: Solvent A (water) and Solvent B
(methanol): 0-20
min 40-100%B (linear gradient), 20-27 min 100%B. HPLC2: Gradient: Solvent A
(water) and
Solvent B (methanol): 0-15 min 40-100%B (linear gradient), 15-25 min 100%B. UV
detection at
' 254nm.
[00263] The compounds of this invention were prepared according to
Figures 1-17.
EXAMPLE 1
SYNTHESIS OF B RING VARIANT COMPOUNDS
B ring variant compounds were synthesized according to Figures 1 and 2.
Oxazole B ring:
Synthesis of (2-Phenyl-oxazol-4-y1)-(3,4,5-trimethoxy-phenyl)-methanone
(36a)iFigure 1):
Ill
CA 3030689 2019-01-18
/\
0 0
0/
0
N 0
411111 (36a)
[00264] (2R)-2-
Phenyl-4,5-dihydro-oxazole-4-carboxylic acid methyl ester (32a).
Acetyl chloride (6.8 mL) was added dropwise to ice-cold methanol (30 mL).
After the addition of
L-serine (0.48 mmol), the reaction mixture was warmed to room temperature (RT)
and stirred
overnight. Evaporation of the solvent gave white solid (2R)-3-hydroxy-2-methyl-
propionic acid
methyl ester HC1 salt, which was used without purification in the next step.
Triethylamine (11
mL, 72.3 mmol) was added slowly to a solution of ethyl benzimidate
hydrochloride (11.6 g, 62.8
mmol) in CH2C12 (150 mL). The reaction mixture was stirred at RT for 30, min
and (2R)-3-
hydroxy-2-methyl-propionic acid methyl ester HCI salt (13.5 g, 79.6 mmol) was
added by
portion. The resulting mixture was stirred for 48 h and concentrated under
reduced pressure. The
compound 32a was separated from flash column as a yellow oil (12.3 g, 95.9%).
NMR
(CDC13) S 7.99 -7.38 (m, 5 H), 4.97 (dd, I H, J = 7.8 Hz, J = 10.5 Hz), 4.70
(t, 1 H, J 8.7 Hz),
4.62 (dd, 1 H, J = 8.7 Hz, J = 10.5 Hz), 3.82 (s, 3 H); MS (ESI) m/z 206.1 (M
+ H)t.
[002651 (2R)-2-
Phenyl-4,5-dihydro-oxazole-4-carboxylic acid (33a). To an ice-cooled
solution of 32a in Me0H/H20 was added LiOH (2.5 equiv) with stirring. The
mixture was
allowed to warm to RT in 1 h, concentrated in vacuo, and the white solid was
dissolved in H20
and acidified with I N HC1 to pH 2.0 and extracted with MgSO4, filtered and
concentrated in
vacuo to provide the acid 33a as a white solid (95.8 %). 111 NMR (CDC13) 8
7.98 (d, 2 H), 7.57-
7.42 (m, 3 H), 5.04 (dd, 1 H, J = 7.8 Hz, J = 10.8 Hz), 4.80 (t, 1 H, J = 8.7
Hz), 4.70 (dd, 1 H, J
= 9.0 Hz, J= 10.8 Hz); MS (ESI) m/z 191.9 (M + H)+, 189.7 (M - 11)-, 145.8 (M -
COOH)..
112
CA 3030689 2019-01-18
[00266] (2R)-2-Phenyl-4,5-dihydro-oxazole-4-carboxylic acid methoxy-
methyl-amide
(34a). To a mixture of 33a (5 mmol), EDO (6 mmol), HOBt (5 mmol) and Et3N (5
mmol) in
CH2Cl2 (50 mL) was added HNCH3OCH3 (5 mmol) and stirring continued at RT for 6-
8 h. The
reaction mixture was diluted with CH2C12 (100 mL) and sequentially washed with
water, satd.
NaHCO3, brine and dried over MgSO4. The solvent was removed under reduced
pressure to yield
a crude product 34a, which was purified by column chromatography as a white
solid (61.0 %). 11-1
NMR (CDCI3) 8 7.98-7.36 (m, 5 H), 7.57-7.42 (m, 3 H), 5.35 (br, t, 1 H), 4.81
(br,. t, 1 H), 4.52
(dd, 1 H, J = 8.7 Hz, J = 10.2 Hz), 3.90 (s, 3 H), 3.27 (s, 3 H); MS (ESI)
rn/z 257.0 (M + H)4.
[00267] (2R)-(2-Ph eny1-4,5 -dihy dro-oxazol-4-y1)-(3,4,5 -trimethoxy-
pheny1)-
methanone (35a). To a solution of n-BuLi (1.6 M, 0.713 mL) in 8 mL THF was
added a solution
of 3,4,5-trimethoxybromobenzene (1.09 mmol) in 3 mL THF under -78 C. The
mixture was
allowed to stir for 2 h and a solution of Weinreb amide 34a (1.14 mmol) in 3
mL THF was
charged. The temperature was allowed to increase at RT and stirred overnight.
The reaction
mixture was quenched with satd. NH4C1, extracted with ethyl ether, dried with
.MgSO4. The
solvent was removed under reduced pressure to yield a crude product, which was
purified by
column chromatography to obtain pure compound 35a as a white solid (47.9 To).
11-1 NMR
(CDCI3) 8 7.97 -7.94 (m, 2 H), 7.62 (s, 2 H), 7.54-7.37 (m, 3 H), 5.61 (q, 1
H, J = 7.5 Hz, 9.9
Hz), 5.12 (t, 1 H, J = 7.5 Hz), 4.57 (q, 1 H, J = 7.8 Hz, 9.9 Hz), 3.96 (s, 6
H), 3.95 (s, 3 H); MS
(ES!) miz 364.1(M + Na)+, 340.1 (M - H).
[00268] (2-Phenyl-oxazol-4-y1)-(3,4,5-trimethoxy-phenyl)-methanone (36a). A
mixture
of 35a (1:48 mmol), CBrCI3 (2.59 mmol) and DBU (2.97 mmol) in CH2Cl2 (20 mL)
was stirred
overnight. The reaction mixture was absorbed on silica gel and purified by
column
chromatography to yield pure 36a as desired (61.6 %). 1H NMR (CDCI3) 8 8.37
(s, 1 H), 8.14-
8.12 (m, 2 H), 7.74 (s, 2 H), 7.52-7.49 (m, 3 H), 3.97 (s, 9 H); MS (ES!) m/z
362.1(M + Na).
[00269] Benzene, pyrimidine, pyridine, furan, thiophene, thiazole, pyrazole
and
= piperidine B ring variants (Figure 2): B ring variants (la-id, lk) were
obtained from their
corresponding acids (37a-37d, 37k). Compound if with thiophene in B ring
position can not be
separated from the mixture of if and a Grignard reagent coupling by-product
3,4,5,3',4',5'-
hexamethoxybiphenyl using flash column. So an alternative method was. used to
prepare if:
Weinreb amide 38f was converted into its corresponding aldehyde which was
further reacted with
3,4,5-trimethoxyphenylmagnesium bromide to afford the alcohol 40f, which can
be easily
113
CA 3030689 2019-01-18
separated from 3,4,5,3',4',5'-hexamethoxybiphenyl using flash column
chromatography.
Oxidation with pyridinium dichromate (PDC) or DMSO did not afford If from
secondary alcohol
40f with good yields. But using Dess-Martin periodinane reagent as oxidant
successfully formed
the desired ketone compound if. le and li were prepared from alcohols 40e and
40i using a
similar method. Compound lg was obtained via a coupling reaction from
piperidine 41g and
3,4,5-trimethox ybenzoic acid.
Benzene B ring:
Synthesis of Biphenyl-3-y1(3,4,5-trimethoxyphenyl)methanone (la) (Figure 2)
OMe
0
40 OMe
OMe
(la)
[00270] N-Methoxy-N-methylbipheny1-3-carboxamide (38a). To a mixture of
37a (5
mmol), EDCI (6 mmol), HOBt (5 mmol) and NMM (11 mmol) in CH2C12 (50 mL) was
added
HNCH30CH3HC1 salt (5 mmol) and stirring continued at RT for 2 h. The reaction
mixture was
diluted with CH2C12. (100 mL) and sequentially washed with water, satd.
NaHCO3, brine and
dried over MgSO4. The solvent was removed under reduced pressure to yield a
colorless oil,
which was used for next step (58.4 %). MS (ESI) rn/z. 264.0 (M + Na).
[00271] Biphenyl-3-y1(3,4,5-trimethoxyphenyl)methanone (la). To a
solution of 38a
(Figure 2) (0.174 g, 0.72 mmoL) in 5 mL THF was added a THF solution of 3,4,5-
trimethoxyphenylmagnesiumbromide (0.5 N, 1.08 mmol) at 0 C. The mixture was
allowed to
stir for 30 min and quenched with satd. NI14C1, extracted with ethyl ether,
dried with MgSO4.
The solvent was removed under reduced pressure to yield a crude product, which
was purified by
column chromatography to obtain pure compound la as a white solid (43.8%). 11-
1 NMR
(CDC13) 8 8.02 (t, 1 H), 7.84-7.74 (m, 2 H), 7.64-7.38 (m, 6 H), 7.11 (s, 2
H), 3.95 (s, 3 H), 3.88
(s, 6 H); MS (ESI) mh 371.1(M + Na).
114
CA 3030689 2019-01-18
Pyrimidine B ring:
Synthesis of (6-Phenylpyrimidin-4-y1)(3,4,5-trimethoxyphenyl)methanone (lb)
(Figure 2)
OMe
0
OMe
N OMe
11101 (lb)
[00272] N-
Methoxy-N-methyl-6-phenylpyrimidine-4-carboxamide (38b). To a mixture
of 37b (5 mmol), EDC1 (6 mmol), HOBt (5 mmol) and NMM (11 mmol) in CH2C12 (50
mL) was
added HNCH30CH3HC1 salt (5 mmol) and stirring continued at RT for overnight.
The reaction
mixture was diluted with CH2C12 (100 mL) and sequentially washed with water,
satd. NaHCO3,
brine and dried over MgSO4. The solvent was removed under reduced pressure to
yield a crude
product, which was purified by column chromatography to obtain pure compound
38b as a
yellow solid (62.3 %). NMR (CDC13) 8 9.28 (s, 1 H), 8.14-8.06 (m, 2 H),
7.96 (br, s, 1 H),
7.54-7.50 (m, 3 H), 5.35 (br, t, 1 H), 4.81 (br, t, 1 H), 4.52 (dd, 1 H, J =
8.7 Hz, J = 10.2 Hz),
3.79 (s, 3 H), 3.42 (s, 3 H); MS (ESI) nVz 266.0 (M + Na)+.
[00273] - (6-
Phenylpyrimidin-4-yI)(3,4,5-trimethoxyphenyl)methanone (lb). To a
solution of 38b (0.243 g, 1 mmoL) in 5 mL THF was added a THF solution of
3,4,5-
trimethoxyphenylmagnesiumbromide (0.5 N, 5.6 mL, 1.4 mmol) at 0 C. The
mixture was
allowed to stir for 30 min and quenched with satd. N1140, extracted with ethyl
ether, dried with
MgSO4. The solvent was removed under reduced pressure to yield a crude
product, which was
purified by column chromatography to obtain pure compound lb (52.3%). Ili NMR
(CDC13)
9.40(d, 1 H,) = 1.5 Hz), 8.29 (d, 11-1, J = 1.5 Hz), 8.22-8.18, 7.57-7.54 (m,
5 H), 7.46 (s, 2 H),
3.96*(s, 3 H), 3.91 (s, 6 H); MS (ES1) mix 351.I(M + H).
115
CA 3030689 2019-01-18
Pyridine B ring:
Synthesis of (6-Phenylpyridin-2-y1)(3,4,5-trimethoxyphenyl)methanone (1c)
(Figure 2)
OMe
0
OMe
I N OMe
(1c)
[00274] N-
Methoxy-N-methyl-6-phenylpicolinamide (38c). To a mixture of 37c (137
mmol), EDCI (2.12 mmol), HOBI (1.86 mmol) and NMM (3.54 mmol) in CH2C12 (20
mL) was
added 1iNCH3OCH3HCI salt (1.86 mmol) and stirring continued at RT for
overnight. The
reaction mixture was diluted with CH2C12 (40 mL) and sequentially washed with
water, said.
NaHCO3, brine and dried over MgSO4. The solvent was removed under reduced
pressure to yield
a crude product, which was purified by column chromatography to obtain pure
compound 38c as
a colorless oil (512 %). NMR (CDCI3) 5 8.02 (d, 1 H, J = 7.0 Hz), 7.86-7.81
(m, 2 H), 7.55
(br, 1 1-1), 7.48 (t, 2 H), 7.44-7.41 (m, 1 H), 3.82 (s, 3 H), 3.44 (s, br, 3
H); MS (ESI) miz 265.0
(M + Na).
[00275] (6-
Phenylpyridin-2-y1)(3,4,5-trimethoxyphenyl)methanone (1c). To a solution
of 38c (0.210g, 0.86 mmoL) in 5 mL THF was added a TI-IF solution of 3,4,5-
trimethoxyphenylmagnesiumbromide (0.5 N, 3.5 mL, 1.73 mmol) at 0 C. The
mixture was
allowed to stir for 30 min and quenched with water, extracted with ethyl
acetate and dried with
MgSO4. The solvent was removed under reduced pressure to yield a crude
product, which was
purified by column chromatography to obtain pure lc as white needle crystals
(78%). NMR
(CDCI3) 8 8.10 (d, br, 2 H), 8.02-8.00 (m, 1 H), 7.97-7.96 (m, 2.H), 7.66 (s,
2 H), 7.49-7.43 (m, 3
II), 3.97 (s, 3 1-1), 3.89 (s, 6 H); MS (ESI) m/z 372.6 (M + Na)+.
116
CA 3030689 2019-01-18
Furan B ring:
Synthesis of (5-Phenylfuran-2-yI)(3,4,5-trimethoxyphenyl)methanone (1d)
(Figure 2)
= OMe
0
OMe
0 OMe
=
(1d) *
[00276] N-Methoxy-N-
methyl-5-phenylfuran-2-carboxamide (38d). To a mixture of
37d (10 mmol), EDCI (12 mmol), HOBt (II mmol) and NMM (21 mmol) in CH2Cl2 (200
mL)
was added liNCH30CH3HCI salt (10.5 mrnol) and stirring continued at RT for
overnight. The
reaction mixture was diluted with CH2Cl2 (200 mL) and sequentially washed with
water, satd.
NaHCO3, brine and dried over MgSO4. The solvent was removed under reduced
pressure to yield
a crude product, which was purified by column chromatography to obtain pure
compound 38d.
(95.2 %). 'H NMR (CDC13) 8 7.82 (d, 1 H, J = 7.0 Hz), 7.46-7.43 (t, 2 H), 7.37-
7.34 (m, 1 H),
7.25 (d, I H, J = 4.0 Hz), 6.78 (d, 1 H, J = 4.0 Hz), 3.86 (s, 3 1-1), 3.41
(s, 3 H); MS (ES!) m/z
254.1 (M + Na).
[00277] (5-
Phenylfuran-2-yI)(3,4,5-trimethoxyphenyl)methanone (1d). To a solution of
38d (0.231 g, 1 rrunoL) in 5 mL THF was added a THF = solution of 3,4,5-
trimethoxyphenylmagnesiumbromide (0.5 N, 4.0 mL, 2 mmol) at 0 C. The mixture
was allowed
to stir for 30 min and quenched with water, extracted with ethyl acetate and
dried with MgSO4.
The solvent was removed under reduced pressure to yield a crude product, which
was purified by
column chromatography to obtain pure compound id as white crystals (35.5%).
NMR
(CDCI3) 8 7.85-7.82 (m, 1 H), 7.48-7.36 (m, 4 H), 7.35 (s, 2 H), 7.25 (d, 1 H,
J = 4:0 Hz), 6.86
(d, 1 H, J = 4.2 Hz), 3.96 (s, 3 H), 3.95 (s, 6 H); MS (ES!) m/z 339.1 (M +
H).
117
CA 3030689 2019-01-18
Thiazole B ring:
Synthesis of (2-Phenylthiaz,o1-5-y1)(3;4,5-trimethoxyphenyl)methanone (le)
(Figure 2)
OMe
0
OMe
N,, S OMe
(le)
[00278] (2-Phenylthiazol-5-y1)(3,4,5-trimethoxyphenyl)methanol (40e).
To a solution of
2-phenylthiazole-5-carbaldehyde 38e (0.567 g, 3 mmoL) in 15 mL-THF was added a
THF
solution of 3,4,5-trimethoxyphenylmagnesiumbromide (0.5 N, 6.5 mL, 3.25 mmol)
at 0 C. The
mixture was allowed to stir for 30 min and quenched with satd. NH4C1,
extracted with ethyl ether:
dried with MgSO4. The solvent was removed under reduced pressuitre to yield a
crude product,
which was purified by column chromatography to obtain pure compound 40e (72.9
%). 1H NMR
(CDC13) S 7.90 (m, 2 H), 7.64 (s, I H), 7.41 (m, 3 H), 6.69 (s, br, 2 H), 6.04
(s, 1 H), 3.86 (s, 6
H), 3.85 (s, 3 H), 1.57 (d, I H, J = 5.5 Hz); MS (ES1) m/z 358.1 (M + Na).
[00279] (2-Phenylthiazol-5-y1)(3,4,5-trimethoxyphenyl)methanone (le).
To a solution
of 40e (0.357 g, 1 mmoL) in 40 thL anhydrous CH2Cl2 was added Dess-Martin
reagent (0.848 g,
2 mmol). The mixture was allowed to stir for 30 mm and quenched with sat.
Na2S203 solution,
extracted with ethyl acetate and dried with MgSO4. The solvent was removed
under reduced
pressure to yield a crude product, which was purified by column chromatography
to give pure
compound le (80.1%). NMR (CDC13) 88.33 (s, 1 H), 8.04 (m, 2 H), 7.51 (m, 3
H), 7.18 (s, 2
H), 3.96 (s, 3 H), 3.93 (s, 611); MS (ESI) rri/z 378.1 (M + H)+.
118
CA 3030689 2019-01-18
Thiophene B ring:
Synthesis of (5-Phenylthiophen-3-y1)(3,4,5-trimethoxyphenyl)methanone (10
(Figure 2)
OMe
0
OMe
S OMe
(10
[002801 N-Methoxy-N-methyl-5-phenylthiophene-3-carboxamide (380. To a
mixture of
37f (2.5 mmol), EDC1 (2.9 mmol), HOBt (2.6 mmol) and NMM (5.3 mmol) in CH2C12
(30 mL)
was added HNCH30CH3HC1 salt (2.6 mmol) and stirring continued at RT for
overnight. The
reaction mixture was diluted with CH2C12 (20 mL) and sequentially washed with
water, satd.
NaHCO3, brine and dried over MgSO4. The solvent was removed under reduced
pressure to yield
a crude product, which was purified by column chromatography to obtain pure
compound 38f.
(90.8 %). 1H NMR (CDC13) 58.28 (d, 1 H, J = 1.5 Hz), 7.69 (d, 1 H, J = 1.5
Hz), 7.64 (d, 2 H, J
= 7.0 Hz), 7.44 (t, 2 H, J = 7.0 Hz), 7.35-7.32 (m, 1 H), 6.78 (d, 1 H, J =
4.0 Hz), 3.86 (s, 3 H),
141 (s, 3 H); MS (ESI) mlz 270.0 (M + Na).
[00281] (5-Phenylthiophen-3-y1)(3,4,5-trimethoxyphenyl)methanol (401).
At -78 C, to
a solution of 381(2.5 mmol) in 5 mL THF under argon protection was added a
solution of LiA1H4
in THF (1 N, 1.42 mL) and stirring continued at 1 h at -20 C. The reaction
mixture was placed
on an ice bath and quenched by 20% H2SO4 solution, extracted with ethyl
acetate and dried over
MgSO4. The solvent was removed under reduced pressure and purified by column
chromatography to yield 5-phenylthiophene-3-carbaldehyde (not shown) (84.8%).
'H NMR
(CDCI3) 59.98 (s, I H), 8.04 (d, 1 H, J = 1.5 Hz), 7.86 (br, 1 H), 7.61-7.58
(br, 2 H), 7.47-7.33
(m, 3 H), 7.35-7.32 (m, 1 H), 6.78 (d, 1 H, J = 4.0 Hz); MS (ESI) ink 210.9 (M
+ Na)*. To a
solution of 5-phenylthiophene-3-carbaldehyde (0.195 g, 1.04 mmoL) in 5 mL THF
was added a
THF solution of 3,4,5-trimethoxyphenylmagnesiumbromide (0.5 N, 2.3 mL, 1.14
mmol) at 0 C.
The mixture was allowed to stir for 30 min and quenched with satd. NH4C1,
extracted with ethyl
ether, dried with MgSO4. The solvent was removed under reduced pressure to
yield a crude
product, which was purified by column chromatography to obtain pure compound
40f.
119
CA 3030689 2019-01-18
(70.5%).11-1 NMR (CDC13) ö 7.55-7.52 (m, 2 H), 7.40-7.35 (m, 3 H), 7.30 (br, 1
H), 7.20 (br, 1
H), 6.72 (s, 2 H), 6.01 (d, 1 H, J = 3.9 Hz), 3.86 (s, 6 H), 3.85 (s, 3 H),
2.42 (d, 1 H, J = 3.9 Hz);
MS (ESI) rn/z 339.1 (M -
[00282] (5-Phenylthiophen-3-y1)(3,4,5-trimethoxyphenyl)methanone (10.
To a solution
of 40f (0.260 g, 0.73 mmoL) in 20 mL anhydrous CH2C12 was added Dess-Martin
reagent (0.465
g, 1.36 mmol). The mixture was allowed to stir for 30 min and quenched with
sat. Na2S201
solution, extracted with ethyl acetate and dried with MgSO4. The solvent was
removed under
reduced pressure to yield a crude product, which was purified by column
chromatography to give
pure compound if as light yellow crystals (60.9%). 1H NMR (CDCI3) ö7.97 (d, 1
H, J = 1.5 Hz),
7.82 (d, 1 H, J = 1.5 Hz), 7.59-7.57 (m, 2 H), 7.45-7.34 (m, 3 H), 7.19 (s, 2
H), 3.95 (s, 3 H),
3.93 (s, 6 H); MS (ES!) m/z 355.1 (M + H).
Piperidine B ring:
Synthesis of (4-Pheny1piperidin-1-y1)(3,4,S-trimethoxyphenypmethanone (1g)
(Figure 2)
OMe
0 ipOMe
OMe
411 (1g)
[00283] (4-Phenylpiperidin-1-y1)(3,4,5-trimethoxyphenyl)nethanone (1g).
To a ,
mixture of 4-phenylpiperidine 41g (5 mmol), EDCI (6 mmol), HOBt (5.5 mmol) and
NMM (6
mmol) in CH2C12 (50 mL) was added 3,4,5-trimethoxybenzoic acid (5.3 mmol) and
stirring
continued at RT for overnight. The reaction mixture was diluted with CH2Cl2
(100 mL) and
sequentially washed with water, satd. NaHCO3, brine and dried over MgSO4. The
solvent was
removed under reduced pressure to yield a crude product, which was purified by
column
chromatography to obtain pure compound lg. (57.9%). Iff NMR (CDCI3) 8 7.35-
7.21 (in, 5 H),
6.66 (s, 2 H), 4.84 (br, 1 H), 3.95 (br, 1 H), 3.88 (s, 6 H), 3.86 (s, 3 H),
3.20-2.87 (br, 2 H), 2.85-
2.74 (tt, 1 H, J = 3.6 Hz, J = 15.6 Hz) 1.92 (br, 2 H), 1.70 (br, 2 H); MS
(ES!) m/z 378.1 (M +
NW.
120
CA 3030689 2019-01-18
Isoxazole B ring:
Synthesis of (5-Phenylisoxazol-3-y1)(3,4,5-trimethoxyphenyl)nethanone (10
(Figure 2)
OMe
0
OMe
N-
0 / OMe
(1i)
[00284] (5-
Phenylisoxazol-3-y1)(3,4,5-trimethoxyphenyl)methanol (40i). To a solution
of 5-phenylisoxazole-3-carbaldehyde 381 (0.365 g, 2.1 mmol) in 15 mL THF was
added a THF
solution of 3,4,5-trimethoxyphenylmagnesiumbromide (0.5 N, 5.5 mL, 2.74 mmol)
at 0 C. The
mixture was allowed to stir for 30 min and quenched with satd. NH4CI,
extracted with ethyl ether,
dried with MgSO4. The solvent was removed under reduced pressure to yield a
crude product,
which was purified by column chromatography to obtain pure compound 401 as a
white solid.
(48.8%).11-1 NMR (CDCI3) 8 7.78-7.77 (m, 2 H), 7.48-7.46 (m, 3 H), 6.74 (s, 2
H), 6.45 (s, 1 H),
5.98 (d, 1 H, J = 3.5 Hz) 3.89 (s, 6 H), 3.86 (s, 3 H), 2.77 (d, 1 H, J = 3.5
Hz); MS (ESL) inh
364.1 (M + Na)+.
[00285] (5-
Phenylisoxazol-3-y1)(3,4,5-trimethoxyphenypmethanone (1i). To a solution
of 40i (0.110 g, 0.73 mmoL) in 8 mL anhydrous CH2Cl2 was added Dess-Martin
reagent (0.274
g, 0.645 mmol). The mixture was allowed to stir for 30 min and quenched with
sat. Na2S203
solution, extracted with ethyl acetate and dried with MgSO4. The solvent was
removed under_
reduced pressure to yield a crude product, which was purified by column
chromatography to give
pure compound 11(70.1%). NMR
(CDC13) 87.87-7.85 (m, 2 H), 7.72 (s, 2 H), 7.53-7.49 (m, 3
H), 7.05 (s, 1 H), 7.82 (d, 1 H, J = 1.5 Hz), 3.97 (s, 3 H), 3.96 (s, 6 H); MS
(ESI) nilz 362.1 (M
+ H)+.
=
121
CA 3030689 2019-01-18
Pvrazole B ring:
Synthesis of (3-Pheny1-1H-pyrazo1-5-y1)(3,4,5-trimethoxyphenyl)methanone (1k)
(Figure
2)
OMe
0
HN OMe
OMe
(1k)
[00286] (3-Phenyl-1H-pyrazol-5-y1)(3,4,5-trimethoxyphenypmethanone (1k) was
prepared
using the same method as used of compound lc from 3-phenyl-1H-pyrazole-5-
carboxylic acid,
IFI NMR (500MHz, CDC130 0 6 10.97 (br, 1 H), 7.77 (s, br, 2 H), 7.48- 7.38 (m,
5 H), 7.14 (s,
br, 1 H), 3.96 (s, 3 II), 3.94 (s, 6 H); MS (ESI) iniz 361.1(M + Na), 337.0 (M
- H.
EXAMPLE 2
SYNTHESIS OF COMPOUNDS OF THIS INVENTION HAVING DIFFERENT Y
LINKERS
[00287] The compounds of this invention possess different Y linkers. Such
compounds, with
different Y linkers, were synthesized according to Figures 3 and 4.
[00288] Compound 1 h was synthesized from 2-phenyl-4,5-dihydro-thiazole-4-
carboxylic acid
42a through three steps described before (Lu, Y.; Wang, Z.; Li, C. M.; Chen,
J.; Dalton, J. T.; Li,
W.; Miller, D. D., Synthesis, in vitro structure-activity relationship, and in
vivo studies of 2-
arylthiazolidine-4-carboxylic acid amides as anticancer agents. Bioorg Med
Chem 2010, 18, (2),
477-95). 1k was converted to oxime isomers 2e-cis,trans and 2f-cis,trans upon
reaction with
hydroxylamines, NH2OH or NH2OCH3. Assignments were made on the basis of
chemical and
spectral data as described infra. An improved Beckmann rearrangement readily
produced the
rearranged amides 2g and 2h from the two geometric stereoisomers 2e-cis and 2e-
trans via their
122
CA 3030689 2019-01-18
1.
reaction with tosyl chloride and subsequent basic aluminum oxide column.
Hydrazide derivatives
2d-cis and 2d-trans were prepared by mixing lh with hydrazine hydrate in
ethanol and refluxing
for 24 h. Acrylonitriles 2c-trans,cis were obtained from Wittig reaction of lh
with diethyl
cyanomethylphosphonate. Cyanoimine 2j was prepared using the procedure as by
described by
Cuccia (Cuccia, S. J.; Fleming, L. B.; France, D. J., A novel and efficient
synthesis of 4-phenyl-
2-chloropyrimidines from acetophenone cyanoimines. Synthetic Communications
2002, 32, (19),
3011-3018). The carbonyl group in compound lh was also reduced to a secondary
alcohol 2b or
converted to an alkene (2a) as illustrated in Figure 3.
[00289] Attempts to remove the carbonyl group between B and C rings in lh,
resulted in the
formation of compound 2i as shown in Figure 4. Introducing cis- and trans-
double bonds into
the carbonyl position formed compounds (3a and 3b), which were synthesized
from a Wittig
reaction with 2-phenylthiazole-4-carbaldehyde. The sulfide compound 4a,
sulfone 4b and
sulfoxide 4c were prepared using 3-aminobiphenyl as starting material through
an initial
Sandmeyer reaction to yield carbonodithioate 52a, followed by CuI catalyzed
coupling reaction
and m-CPBA oxidation. Sulfonamide linked compound 4d was prepared from
reaction of 3-
biphenylsulfonyl chloride with 3,4,5-trimethoxyaniline in the presence of NEt3
in DMF.
Synthesis of (2-Phenyl-thiazol-4-y1)-(3,4,5-trimethoxy-phenyl)-methanone (1h)
[Figure 3]
OMe
0
OMe
S N OMe
(1h)
[00290] (2-Phenyl-thiazol-4-y1)-(3,4,5-trimethoxy-phenyl)-methanone (1h). A
mixture of 2-
pheny1-4,5-dihydrothiazole-4-carboxylic acid (5 mmol), EDCI (6 mmol) and HOBt
(5 mmol) in
CH2C12 (50 mL) was stirred for 10 min. To this solution, NMM (5 mmol) and
123
CA 3030689 2019-01-18
HNCH3OCH3 (5 mmol) were added and stirring continued at RT for 6-8 h. The
reaction mixture
was diluted with CH2C12 (100 mL) and sequentially washed with water, satd.
NaHCO3, brine and
dried over MgSO4. The solvent was removed under reduced pressure to yield a
crude product,
which was purified by column chromatography to get 2-phenyl-4,5-
dihydrothiazole-4-carboxylic
123a
CA 3030689 2019-01-18
acid methoxymethylamide. A solution of 2-phenyl-4,5-dihydrothiazole-4-
carboxylic acid
methoxymethyrarnide (1 equiv) in CH2C12 was cooled to 0 C, and distilled DBU
(2 equiv) was
added. Bromotrichloromethane (1.7 equiv) was then introduced dropwise via
syringe over 10
min. The reaction mixtures were allowed to warm to RT and stirred overnight.
Upon washing
with satd. aqueous NH4C1 (2 x 50 mL), the aqueous phase was extracted with
Et0Ac (3x 50 mL).
The combined organic layers were dried on MgSO4, filtered and concentrated in
vacuo. The
residue was purified by flash chromatography as needed providing 2-phenyl-
thiazole-4-
carboxylic acid methoxymethylamide (73.6 %). NMR
(300MHz, CDC13) 8.01 (s, 1 H), 7.99-
7.96 (m, 2 H), 7.47-7.44 (m, 3 H), 3.88 (s, 3 H), 3.49 (s, 3 H). MS (ESI) miz
271.0 (M + Na).
To a solution of 3,4,5-trimethoxyphenylmagnesium bromide (0.5 N, 3 mL) in 2 mL
THF was
charged a solution of 2-phenyl-thiazole-4-carboxylic acid methoxymethylamide
(1 mmol) in 3
mL THF at 0 C. The mixtures were stirred for 30 min until amides disappeared
on TLC plates.
The reaction mixture was quenched with satd. NR4C1, extracted with ethyl
ether, dried with
MgSO4. The solvent was removed under reduced pressure to yield a crude
product, which was
purified by column chromatography to obtain pure compound lb. Yield: 27.3 %.
11-1 NMR (300
MHz, CDC13) 5 8.29 (s, 1 H), 8.03 (q, 2 H), 7.80 (s, 2 H), 7.49-7.47 (m, 3 H),
3.96 (s, 6 H), 3.97
(s, 3 H). MS (ES I) m/z 378.1 (M + Na).
Synthesis of 4-(2-Methy1-1-(3,4,5-trimethoxyphenyl)prop-1-eny1)-2-
phenylthiazole (2a)
[Figure 31
OMe
OMe
S r N OMe
(2a)
[00291] 4-(2-Methy1-1-(3,4,5-trimethoxyphenyl)prop-1-enyl)-2-
phenylthiazole (2a)
[Figure 31. At -78 C, to a solution of 223 mg isopropyl triphenylphosphonium
iodide (0.52
124
CA 3030689 2019-01-18
mmol) in 5 mL of THF was added dropwise 0.4 mL of 1.6 N n-BuLi in. hexane
under Ar2
protection. And the mixture was stirred at 0 C for 40 min. A solution of 140
mg (0.39 mmol) of
lb in 5 mL of THF was added dropwise at 0 C, and the mixture was stirred for
1 h at RT. The
reaction mixture was treated with saturated NH4C1 solution. After a
conventional workup,
column chromatography (silica gel, petroleum ether/ethyl acetate) gave
compound 2a (86 mg,
57.3 %). NiVIR
(300 MHz, CDC13) 5 7.98-7.97 (m, 2 H), 7.45-7.40 (m, 3 H), 6.77 (s, 1 H),
6.48 (s, 2 H), 3.86 (s, 3 H), 3.82 (s, 6 H), 2.15 (s, 3 H), 1.81 (s, 3 H). MS
(ESI) m/z 404.1 (M +
Na)'.
Synthesis of (2-Phenylthiazol-4-y1)(3,4,5-trimethoxyphenyl)methanol
(2b)[Figure 3]
OMe
HO
OMe
s N OMe
410 (2b):
[00292] 2-
Phenyl-4,5-dihydrothiazole-4-carboxylic acid (42a). Benzonitrile (40 mmol)
was combined with L-cysteine (45 mmol) in 100 mL of 1:1 Me0H/pH 6.4 phosphate
buffer
solution. The reaction was stirred at 40 C for 3 days. The precipitate was
removed by filtration,
and Me0H was removed using rotary evaporation. To the remaining solution was
added 1M HCl
to adjust to pH = 2 under 0 C. The resulting precipitate was filtered to
yield a white solid 2-
pheny1-4,5-dihydrothiazole-4-carboxylic acid 42a, which was used directly to
next step without
purification.
[00293] 2-
Phenylthiazole-4-carbaldehyde (42b). At -78 C, to a solution of 2-phenyl-
thiazole-4-carboxylic acid methoxymethylamide (lequiv) in THF was added LiA1H4
(1 equiv, 1
N in THF) and stirring for 1 h at -20 C. The reaction mixture was placed on
an ice bath and
quenched by 20% H2SO4 solution, extracted with ethyl acetate and dried over
MgSO4. The
solvent was removed under reduced pressure and purified by column
chromatography to yield
42b (45.8 %). NMR
(300 MHz, CDC13) 5 10.1 (s, 1 H), 8.17 (s, 1 H), 8.02-8.00 (m, 2 H),
7.50-7.48 (m, 3 H). MS (ESI) m/z 244.1 (M -F Na + Me0H)t.
125
CA 3030689 2019-01-18
[00294] (2-Phenylthiazol-4-y1)(3,4,5-trimethoxyphenypmethanol (2b)
[Figure 3). At 0
C, to a solution of 104 mg of 42b (0.55 mmol, 1 eq.) in 6 mL THF was added
3,4,5-
trimethoxyphenylmagnesium bromide (0.5 N in THF, 2.9 mL). The mixtures were
stirred for 30
min until aldehyde disappeared on TLC plates. The reaction mixture was
quenched with satd.
NH4CI, extracted with ethyl ether, dried with MgSO4. The solvent was removed
under reduced
pressure to yield a crude product, which was purified by column chromatography
to obtain pure
compound (2b). 11-1 NMR (300 MHz, CDC13) 8 7.95-7.92 (m, 2 H), 7.44-7.43 (m, 4
H), 6.97 (s, I
H), 6.76 (s, 2 H), 5.93 (d, 1 H, J = 3.6 Hz), 3.86 (s, 9 H). MS (ES!) tri/z
402.1 (M + Na).
Synthesis of (Z)-3-(2-phenylthiazol-4-y1)-3-(3,4,5-
trimethoxyphenyl)acrylonitrile (2c-trans)
and (E)-3-(2-phenylthiazol-4-y1)-3-(3,4,5-trimethoxyphenyl)acrylonitrile (2c-
cis) [Figure 3]
CN
OMe OMe
NC \
OMe OMe =
S N OMe S N OMe
411 4111
(2c-cis) (2c-trans)
(Z)-3-(2-phenylthiazol-4-y1)-3-(3,4,5-trimetboxyphenyl)acrylonitrile (2c-
trans). To a solution
of 0.4 mL of 2.5 N n-BuLi in hexane and 10 mL of THF was added dropwise a
solution of 177
mg (1 mmol) of diethyl cyanomethylphosphonate in 5 mL of THF at 0 C under Ar2.
The ice bath
was removed, and the mixture was stirred at 25 C for 40 min. A solution of
200 mg (0.56 mmol)
of lh in 10 mL of THF was added dropwise at 0 C, and the mixture was stirred
for 1 h at RT.
The reaction mixture was treated with saturated NH4C1 solution. After a
conventional workup,
column chromatography (silica gel, petroleum ether/ethyl acetate) gave
compounds 2c-trans (83
mg) and 2c-cis (76 mg). IF1 NMR (300 MHz, CDCI3) 5 8.01-7.99 (m, 2 H), 7.44-
7.40 (m, 3 H),
7..21 (s, 1 H), 6.74 (s, 2 H), 6.67 (s, 1 H), 3.93 (s, 3 H), 3.89 (s, 6 H). MS
(ES!) rri/z 401.1 (M +
Na).
126
CA 3030689 2019-01-18
[00295] (E)-3-
(2-phenylthiazol-4-y1)-3-(3,4,5-trimethoxyphenyflacrylonitrile (2c-cis).
11-1 NMR (300 MHz, CDC13) 8. 8.07-8.05 (m, 2 H), 7.49-7.46 (m, 4 H), 6.66 (s,
2 H), 5.64 (s, 1
H), 3.91 (s, 3 H), 3.86 (s, 6 H). MS (ESI)m/z 401.1 (M + Na).
Synthesis of (Z)-4-(hydrazono(3,4,5-trimethoxyphenyl)methyl)-2-phenylthiazole
(2d-cis)
and (E)-4-(hydrazono(3,4,5-trimethoxyphenyl)methyl)-2-phenylthiazole (2d-
trans) [Figure
3]
NH2 OMe
OMe
H2N N
OMe OMe
S N OMe S .Aµ1 OMe
110
(2d-cis) (2d-trans)
[00296] (Z)-4-
(hydrazono(3,4,5-trimethoxyphenypmethyl)-2-phenylthiazole (2d-cis).
To a mixture of lh (230 mg, 0.65 mmol) in 3 mL CH2C12 and 3 mL ethanol was
added hydrazine
hydrate (2 mL). Then the mixture was refluxed for overnight. After completion
of the reaction,
the residue was absorbed on silica gel and purified by column chromatography
to give
compounds 2d-cis (80 mg) and 2d-trans (56 mg). 1H NMR (300 MHz, CDC13) 8 8.01-
7.98 (m, 2
H), 7.49-7.46 (m, 5 H), 7.33 (s, 1 H), 6.82 (s, 2 H), 3.87 (s, 3 H), 3.85 (s,
6 H). MS (ESI) nilz
370.1 (M + H).
[00297] (E)-4-(hydrazono(3,4,5-trimethoxyphenyl)methyl)-2-phenylthiazole (2d-
trans). Ili NMR (300 MHz, CDC13) 5 8.04-8.01 (m, 2 H), 7.44-7.40 (m, 3 H),
6.95 (s, 1 H), 6.65
(s, 2 H), 5.62 (s, 2 H), 3.93 (s, 3 H), 3.87 (s, 6 H). MS (ES1)m/z 370.1 (M +
H).
127
CA 3030689 2019-01-18
Synthesis of (Z)-(2-Phenylthiazol-4-y1)(3,4,5-trimethoxyphenypmethanone oxime
(2e-cis)
and (E)-(2-Phenylthiazol-4-y1)(3,4,5-trimethoxyphenyl)methanone oxime (2e-
trans) [Figure
3]
OMe OH OMe
HO-N
OMe
\ OMe
S N OMe S ,N OMe
1110 110
(2e-cis) (2e-trans)
[00298] (Z)-(2-
Phenylthiazol-4-y1)(3A5-trimethoxyphenyl)methanone oxime (2e-cis)
To a suspension of lh (210 mg, 0.59 mmol) in 10 mL ethanol was -added. an
aqueous solution (2
mL) of hydroxylamine hydrochloride (127 mg, 1.83 mmol). Then 2 mL 1 N NaOH was
added
dropwise to the reaction mixture and the mixture was stirred at 55 C for 3 h.
After completion of
the reaction, the residue was absorbed on silica gel and purified by column
chromatography to
give compounds 2e-cis (85 mg) and 2e-trans (50 mg). III NMR (300 MHz, DMSO-d6)
8 11.95
(s, 1 H), 8.35 (s, 1 H), 7.91-7.89 (m, 2 H), 7.50-7.44 (br, 3H), 6.85 (s, 2
H), 3.73 (s, 6 H), 3.70 (s,
3.H). MS (ESI)miz 393.1 (M + Na); 368.9 (M ¨ H).
[00299] (E)-(2-Phenylthiazol-4-y1)(3,4,5-trimethoxyphenyOmethanone oxime 2e-
trans). 11-1 NMR (300 MHz, DMSO-d6) 6 11.49 (s, I H), 7.92-7.89 (m, 2 H), 7.64
(s, 1 H), 7.51-
7.49 (m, 3 H), 7.34 (s, 1 H), 6.75 (s, 2 H), 3.75 (s, 6 H), 3.72 (s, 3 H). MS
(ES!) miz 393.1 (M +
Na)+; 368.9 (M ¨ H).
Synthesis of (Z)-(2-Phenylthiazol-4-y1)(3,4,5-trimethoxyphenyl)methanone 0-
methyl
oxime (2f-cis) and (E)-(2-Phenylthiazol-4-y1)(3,4,5-trimethoxyphenyl)methanone
0-methyl
oxime (2f-trans) [Figure 3]
128
CA 3030689 2019-01-18
OMe
Me0¨N pMe OMe
OMe OMe
S N OMe s N OMe
101
(21-cis) (21-trans)
[00300] (Z)-(2-
Phenylthiazol-4-y1)(3,4,5-trimethoxyphenyl)methanone 0-methyl
oxime (2f-cis). To a suspension of lb (110 mg, 0.59 mmol) in 10 mL pyridine
was added 0-
methylhydroxylamine hydrochloride (52 mg, 0.63 mmol) and the mixture was
stirred at 60 C for
overnight. The reaction was quenched with 1 N HCI solution, extracted with
ethyl acetate and
dried with MgSO4. The solvent was removed under reduced pressure to yield a
crude product,
which was purified by column chromatography to give pure compounds 2f-cis (41
mg) and 2f-
trans (33 mg). 11-1 NMR (500 MHz, CDC13) 8 8.13 (s, 1 H), 7.96-7.94 (m, 211),
7.45-7.44 (m, 3
H), 6.94 (s, 2 H), 4.13 (s, 3 H), 3.91 (s, 6 H), 3.88 (s, 3 H). MS (ESI)miz
407.2 (M + Nar.
[00301] (E)-(2-
Phenylthiazol-4-y1)(3,4,5-trimethoxyphenyl)methanone 0-methyl
oxime (2f-trans). Ili NMR (500 MHz, CDCI3) 5 8.00-7.98 (m, 2 H), 7.44-7.43 (m,
3 H), 7.28 (s,
1 H), 6.70 (s, 2 H), 4.08 (s, 3 H), 3.91 (s, 6 H), 3.85 (s, 3 H). MS (ESI) tez
407.0 (M + Na)4.
Synthesis of 2-Phenyl-N-(3,4,5-trimethoxyphenyl)thiazole-4-carboxamide (2g)
[Figure 3]
0
S N OMe
oile0 OMe
(2g)
[00302] 2-Phenyl-N-(3,4,5-
trimethoxyphenyl)thiazo1e-4-carboxamide (2g). To a
solution of 2e-cis (21 mg, 0.06 mmol) in 5 mL CH2C12 was added p-
toluenesulfonyl chloride (23
129
CA 3030689 2019-01-18
mg, 0.12 mmol) and NaH (5 mg, 60% in light mineral oil). Then the reaction
mixture was stirred
for 20 min. After completion of the reaction, the residue was absorbed on
silica gel and purified
by A1203 column chromatography to give compound 2g (15 mg). tH NMR (300 MHz,
CDC13) 5
9.22 (s, 1H), 8.19 (s, 1 H), 8.02-7.99 (m, 2 H), 7.52-7.50 (m, 3 H), 7.07 (s,
2 H), 3.92 (s, 6 H),
3.85 (s, 3 H). MS (ESI) m/z 371.1 (M + Hr.
Synthesis of 3,4,5-Trimethoxy-/V-(2-phenylthiazol-4-yl)benzamide (2h) [Figure
31
0
HN
s N OMe
*le OMe
(2h)
[00303] 3,4,5-Trimethoxy-N-(2-phenylthiazol-4-yl)benzainide (2h). To a
solution of 2e-
trans (26 mg, 0.07 mmol) in 5 mL CH2C12 was added p-toluenesulfonyl chloride
(27 mg, 0.14
mmol) and NaH (5 mg, 60% in light mineral oil). Then the reaction mixture was
stirred for 20
min. After completion of the reaction, the residue was absorbed on silica gel
and purified by
A1203 column chromatography to give compound 2h (15 mg). 11-1 NMR (300 MHz,
CDC13) 5
8.88 (s, 1H), 7.94-7.91 (m, 2 H), 7.83 (s, 1 H), 7.48-7.46 (m, 3 H), 7.18 (s,
2 H), 3.97 (s, 6 H),
3.94 (s, 3 H). MS (ESI) m/z 393.1 (M + Na).
Synthesis of N-4(2-phenylthiazol-4-y1)(3,4,5-
trimethoxyphenyl)methylene)eyanamide (2j)
[Figure 3]
NC
OMe
OMe
S õ N OMe
lio (2j)
130
CA 3030689 2019-01-18
[00304] N42-phenyltbiazol-4-y1)(3,4,5-trimethoxyphenyl)methylene)eyanamide
(2j). 100
mg of lb (0.28 mmol, 1 eq.) was dissolved in 10 mL methylene chloride.
Titanium tetrachloride
in methylene chloride (1.0 N, 0.7 mL, 2.5 eq.) was added dropwise at 0 C and
stirred for 30 min.
Bis-trimethylsilylcarbodiimide (2.4 eq.) in 2 mL methylene chloride was added
and the reaction
stirred overnight protected from air and moisture. The reaction was treated
with ice-water
mixture followed by extraction with methylene chloride. The organic phase was
dried over
magnesium sulfate, filtered through celiteTM and concentrated to give the
crude acetophenone
cyanoimines which were purified by flash column as isomers with a ratio of
3:7. 1H NMR (300
MHz, CDCI3) 8 8.72 (br, 0.3 H), 8.63 (s, 0.7 H), 8.09-8.07 (m, 1.4 H), 7.99
(br, 0.6 H), 7.58-7.56
(br, 3 H), 7.26 (s, 1.4 H), 7.18 (s, 0.6 H), 3.84, 3.83 (s, s, 611), 3.82 (s,
3 H). MS (ESI) mtz 402.1
(M + Na).
Synthesis of N4(4-hydroxy-3,5-dim ethoxyph enyl)(2-ph enylthiazol-4-
yl)methylenc)eyanamide (32).
NC
OMe
OH
S N OMe
1110 (32)
N-44-hydroxy-3,5-dimethoxyphenyl)(2-phenylthiazol-4-yOmethylene)eyanamide (32)
was
obtained as a by-product from synthesis of 2j. 11-1 NMR (500MHz, CDC13 LIII U
8.23 (s, 1 H),
8.02 (m, 2 H), 7.92 (s, 2 H), 7.55 (m, 3 H), 6.02 (s, 1 H), 3.99 (s, 6 H). MS
(ESI) miz 364.1(M +
H)+.
Synthesis of (Z)-2-Phenyl-4-(3,4,5-trimethoxystyryl)thiazole (3a) and (E)-2-
Pheny14-(3,4,5-
trimethoxystyryl)thiazole (3b) [Figure 4]
131
CA 2809256 2017-12-19
CA 3030689 2019-01-18
OMe Me0 OMe
Me0
OMe
Me
S N
---"N
(3a) (3b)
[00305] (Z)-2-
Pheny1-443,4,5-trimethoxystyryl)thiazole (3a). Triphenylphosphine (3.41
g, 13 mmol) was added to a solution of 5-(bromomethyl)-1,2,3-tnimethoxybenzene
(2.61 g, 10
mmol) in dry THF (30 mL). The mixture was refluxed with stirring for 6 h. The
resulting white
solid was filtered and washed with ether/hexane to afford the product 3,4,5-
trimethoxybenzyltriphenylphosphonium bromide in 96.4% yield. ill NMR (500 MHz,
CDCI3) 8
7.77-7.73, 7.65-7.61 (m, 15 H), 6.44 (d, 2 H, J 1.5 Hz), 5.37 (d, 2 H, J = 14
Hz), 3.76 (s, 3 H),
3.51 (d, 6 H); MS (ES!) m/z 443.1 (M - Brr. At -78 C, n-BuLi (0.42 mL, 2.5.N
in hexane) was
added to a solution of 3,4,5-trimethoxybenzyltriphenylphosphonium bromide (500
mg, 0.96
mmol) in 10 triL THF. After stirring at RT for 2 h, aldehyde 42b (109 mg, 0.58
mmol) in 3 mL .
THF was charged and stirred for 30 min. The reaction mixture was treated with
saturated NH4CI
solution. After a conventional workup, column chromatography (silica gel,
petroleum ether/ethyl
acetate) gave compounds 3a (57 mg) and 3b (99 mg). NMR,
(500 MHz, CDC13) 8 7.90-7.89
(m, 2 H), 7.42-7.40 (m, 3 H), 7.07 (s, 1 H), 6.71 (s, 2 H), 6.66 (s, 1 H),
3.87 (s, 6 H), 3.75 (s, 3
II); MS (ES!) m/z 376.1 (M + NW.
[00306] (E)-2-
Phenyl-4-(3,4,5-trimethoxystyryl)thiazole (3b). iff NMR (500 MHz,
CDC13) 88.03-8.01 (m, 2 H), 7.52 (d, 1 H, J = 16 Hz), 7.47-7.44 (m, 3 H), 7.16
(s, 1. H), 7.05 (d,
1 H, J = 16 Hz), 6.79 (s, 2 H), 3.92 (s, 6 H), 3.88 (s, 3 H). MS (ES!) m/z
354.1 (M + H)4.
132
CA 3030689 2019-01-18
Synthesis of Bipheny1-3-y1(3,4,5-trimethoxyphenypsulfane (4a),
3-(3,4,5-
Trimethoxyphenybutfonyl)biphenyl (4b) and 3-(3,4,5-
Trimethoxyphenylsulfinyl)biphenyl
(4c) [Figure 4]
0
OMe OMe
ome
II
OMe 111 OMe
ome
OMe OMe
ome
(4a) (4b) (4c)
[00307] S-
Biphenyl-3-y1 0-ethyl carbonodithioate (52a). To a solution of 1 equiv. of
biphenyl-3-amine (1 g, 5.92 mmol) in water (7.3 mL) at 0 C was added
concentrated
hydrochloric acid (1 mL). A cold solution of 1.1 equiv. of sodium nitrite (450
mg, 6.5 mmol) in
water (3 mL) was added slowly and stirred for 15 min. The cold diazonium
solution was added
slowly to a solution of 1.3 equiv. of potassium ethyl xanthate (1.16 g, 1.3
mmol) in water (1.3
mL) at 45 C. The reaction mixture was stirred for an additional 30 mm at 45
C and then cooled
to RT. The reaction mixture was extracted with diethyl ether (3 x 50 mL). The
combined organic
extracts were washed with 1 N NaOH solution (100 mL), water (3 x 50 mL), brine
(50 mL), dried
over MgSO4, filtered and evaporated under reduced pressure. The resulting
crude xanthate 52a
was used directly in the next step without further purification. MS (ESI) m/z
275.0 (M + H)4.
[00308]
Biphenyl-3-y1(3,4,5-trimethoxyphenyl)sulfane (4a). To a solution of 52a (1.1
g,
crude compound) in ethanol (8 mL) was added potassium hydroxide (2.1 g, 12 mL)
and heated
to reflux for overnight. The solution was cooled to RT and the ethanol was
evaporated under
reduced pressure. The residue was dissolved in water and washed with diethyl
ether (10 mL). The
aqueous layer was acidified with 2 N HC1 and extracted with diethyl ether (3 k
50 mL). The
organic extracts were washed with water (50 mL), brine (50 mL), dried over
MgSO4, filtered and
evaporated under reduced pressure to afford 0.85 g (77.3 %) of crude biphenyl-
3-thiol product
(overall, 3 steps). Into a round-bottomed flask, stirred magnetically, were
placed 0.1 g (1.04
mmol) of sodium tert-butoxide and 83 mg of copper iodide (0.43 mmol). After
the reaction vessel
was sealed, 0.13 g (0.71 mmol) of 4-methoxybenzenethiol and 0.19 g (0.65 mmol)
of 5-iodo-
1,2,3-trimethoxybenzene in 3.0 mL of toluene were injected through the septum.
The reaction
133
CA 3030689 2019-01-18
mixture was heated for overnight at 110 C. Purification was performed by flash
chromatography,
and an amorphous solid was obtained (40% yield). IHNMR (500 MHz, CDC13) 5 7.54-
7.52 (m, 3
H), 7.44-7.41 (in, 3 H), 7.37-7.33 (m, 2 H), 7.23 (s, br, 1 El), 6.69 (s, 2
H), 3.86 (s, 3 H), 3.80 (s, 6
H). MS (ES 1) Trz/z 353.2 (M + H)4.
[00309] 3-(3,4,5-Trimethoxyphenylsulfonyl)biphenyl (4b). To a solution of
60 mg (0.17
mmol) of compound 4a and 5 rriL of dichloromethane was added very slowly 2
equiv. of m-
_CPBA over 3 h. Sulfoxide formation was monitored by thin-layer
chromatography. Purification
was performed with a flash chromatographic column, and an amorphous powder of
(4b) was
obtained (73% yield). 114 NMR (500 MHz, CDC13) 68.14 (br, 1 H), 7.89 (d, 1 H),
7.78 (d, 1 H),
7.59-7.56 (in. 3 Fe, 7.49-7.39 (m, 3 H), 7.19 (s, 2 H), 3.89 (s, 6 H), 3.87
(s, 3 H). MS (ES!) m/z
385.0 (M + Na)+.
[00310] 3-(3,4,5-Trimethoxyphenylsulfinyl)biphenyl (4c). At 0 C, to a
solution of 500
mg (1.42 mmol) of compound (4a) and 5 mL of dichloromethane was added very
slowly 1 equiv.
of m-CPBA over 3 h. Sulfoxide formation was monitored by thin-layer
chromatography.
Purification was performed with a flash chromatographic column, and an
amorphous powder of
(4c) was obtained (87% yield). Ili NMR (500 MHz, CDC13) 67.92 (br, 1 H), 7.71
(d, 2.H), 7.62-
7.60 (in, 3 H), 7.58-7.40 (m, 4 H), 6.94 (s, 2 H), 3.79 (s, 3 H), 3.74 (s, 6
H). MS (ES!) m/z 369.1
(M + H)4.
Synthesis of N-(3,4,5-trimethoxyphenyl)bipheny1-3-sulfonamide (4d) [Figure 4]
0 OCH3
0
.s" OCH3
H
OCH3
[00311] N-(3,4,5-Trimethoxyphenyl)bipheny1-3-sulfonamide (4d). A
mixture of 65 mg
of biphenyl-3-sulfonyl chloride (0.25 mmol), 44 mg of 3,4,5-trimethoxyaniline
(0.24 mmol), and
0.3 mmol of triethylamine in 5 mL DMF 'was stirred overnight. The reaction
mixture was treated
with water and extracted with ethyl acetate. After a conventional workup,
column
chromatography (silica gel, petroleum ether/ethyl acetate) gave 88 mg
compounds (4d) (91.7%).
134
CA 3030689 2019-01-18
NMR (500 MHz, CDC13) 5 7.96 (t, H, J= 1.8 Hz), 7.81-7.74 (in, 2 H), 7.57-7.40
(n, 6 H),
6.33 (s, 2 H), 3.86 (s, 3 H), 180 (s, 6 H). MS (ESI) m/z 422.1 (M + Na).
2-Pheny1-4-(3,4,5-trimethoxyphenypthiazole (2i) [Figure 4]
Me0 OMe
OMe
s N
[00312] 2-Phenyl-4-(3,4,5-trimethoxyphenyOthiazole (2i). Bromine (160
mg, 1 mmol)
was added dropwise to a stirred solution of an 1-(3,4,5-
trimethoxyphenyl)ethanone (210 mg, 1
mmol) in ethanol (30 mL) and the solution was stirred at 0 C for 1 h and then
poured into water
to form a precipitate. This was recrystallized from ethanol to give broi-
noacetophenone (70%) and
used directly for next step. A mixture of bromoacetophenone (288 mg, 1. mmol)
and
benzothioamide (137 mg, 1 mmol) in ethanol was refluxed for 1 h. The reaction
mixture was
concentrated in vacuo and purified with flash column to give 21(167 mg,
51.1%). 11-1 NMR (500
MHz, CDCI3) 5 8.05-8.03 (m, 2 H), 7.48-7.44 (m, 3 H), 7.41 (s, I H), 7.22 (s,
2 H), 3.97 (s, 6 H),
3.89 (s, 3 H). MS (ESI) m/z 350.1 (M + Na).
EXAMPLE 3
SYNTHESIS OF METHOXY BENZOYL THIAZOLE COMPOUNDS HAVING
DIFFERENT "A" RINGS AND/OR SUBSTITUTED "A" RING
[00313] The compounds of this invention possess different substituted
or unsubstituted A
rings such as phenyl or indolyl. Such compounds were synthesized according to
Figures 5 and 6.
[00314] Hydroxyl arid aminomethyl were introduced at the para-position
of the phenyl A-
ring, as well as the phenyl was replaced with 5-indoly1 and 2-indoly1 rings.
Weinreb amides 57a,
61a, 65a, and 67a were prepared by the procedure presented in Figure 5 using
aryl nitriles as
starting materials. 2-Cyano-indole 60a was prepared according to a standard
procedure (Pletnev,
135
CA 3030689 2019-01-18
A. A.; Tian, Q.; Larock, R. C., Carbopalladation of nitrites: synthesis of 2,3-
diarylindenones and
polycyclic aromatic ketones by the Pd-catalyzed annulation of alkynes and
bicyclic alkenes by 2-
iodoarenenitriles. J Org Chem 2002, 67(26), 9276-87). Protections of hydroxyl
(TBDMSC1),
indolyl (PhS02C1) and amino (Boc20) groups were used in preparations.
Deprotection of
TBDMS and oxidation from thiazoline (58a) to thiazole (21) took place in one-
step using
TBAF/THF solution. This thiazoline-thiazole oxidation takes place
spontaneously in the
reaction of thiazoline Weinreb amide and Grignard reagent. The same phenomena
is observed
during preparation of the indole compounds 62a and 66a.
[00315] Compound 62a was separated as a pure thiazole compound after reaction
with 3,4,5-
trimethoxphenyllithium without the need for further oxidation. Compound 66a
was obtained by
removing the phenylsulfonyl protecting groups in hot NaOH ethanol solution.
para-OH and NH2
on the A ring of 21 and 2r were obtained by similar Grignard reactions from
the Weinreb amides
58a and 68a. Compound 2r was further converted to the HCI salt (2r-HCI) and
the HCI salt of
monomethyl amine 2s-HCI using NaH/Mel conditions and dimethylamine 2u under
HCHO/NaBH3CN conditions.
Substituted A ring:
Synthesis of (2-(4-Hydroxyphenyl)thiazol-4-y1)(3,4,5-
trimethoxypheny1)methanone (21)
[Figure 51
OMe
0
OMe
S,N OMe
OH (21)
136
CA 3030689 2019-01-18
1.
(R)-2-(4-Hydroxypheny1)-N-methoxy-N-methy1-4,5-dihydrothiazole-4-carboxamide
(57a)
was synthesized using the same method as used for 38d. Quantitative yield.
NMR (500
MHz, CDC13) 0 7.56 (d, 2 H, J= 8.5 Hz), 6.84 (br, 1 H), 6.73 (d, 2 H, J-
136a
CA 3030689 2019-01-18
8.5 Hz), 5.64 (t, br, 1 H), 3.87 (s, 3 H), 3.30 (s, 3 H). MS (ESI) m/z 289.0
(M + Na), 264.9 (M -
H).
[00317] (R)-(2-(4-(tert-Butyldimethylsi lyloxy)pheny1)-4,5-dihydrothiaz
01-4-y1)(3,4,5 -
trimethoxyphenyl)methanone (58a) was synthesized using the same method as used
for
(35a)-see Example 1. 67.0% yield. 11-1 NMR (300 MHz, CDC13) 8 7.73 (d, 2 11, J
= 8.7 Hz), 7.61
(s, 2 H), 6.83 (d, 2 H,.! = 8.7 Hz), 5.95 (dd, 1 H, J = 8.1 Hz, 9.0 Hz), 4.09,
(d0, 1 H, J = 7.8 Hz,
11.1 Hz), 3.95 (s, 3 H), 3.94 (s, 6 H), 3.55 (dd, 1 H, J = 9.3 Hz, 11.1 Hz),
0.97 (s, 9 H), 0.19 (s, 6
H). MS (ESI) m/z 510.4 (M + Isla)+, 486.0 (M
[00318] (2-(4-Hydroxyphenyl)thiazol-4-y1)(3,4,5-
trimethoxyphenyl)methanone (21). At
0 C, to a solution of 58a (0.2 mmol) in 5 mL CH2Cl2 was added a solution of
tetrabutylammonium fluoride in THF (1 N, 0.6 mmol) and stirred at RT for
around 14 h until
reaction was finished by TLC monitor. 67.0% yield. 11-1 NMR (500 MHz, DMSO-d6)
8 10.1
(s, 1 H), 8.51 (s, 1 H), 7.85 (d, 2 H, J = 8.50 Hz), 7.62 (s, 2 H), 6.91 (d, 2
H, J = 8.5 Hz), 3.86 (s,
6 H), 3.79 (s, 3 H). MS (ESI) ink 394.1 (M + Na)4, 369.9 (M
(2-(4-(Aminomethyl)phenyl)thiazol-4-y1)(3,4,5-trimethoxyphenyl)methanone
hydrochloride (2r or 2r-HC1) [Figure 5]
OMe
0
OMe
S OMe
14111
NH2HCI (2r-HC1)
[00319] (R)-tert-Butyl 4-(4-(methoxy(methyl)carbamoy1)-4,5-
dihydrothiazol-2-
yl)benzyl carbamate (67a). 4-(Aminomethyl)benzonitrile (25.09 g, 0.149 mol)
and L-cysteine
(18.1 g, 0.149 mol) were suspended in 500 mL Me0H and pH 6.4 buffer solutions
(1:1) and
stirred for 3 days at RT. Triethylamine (30 mL) was added to the mixture and
Boc20 (68 g,
0.31mo1) was added to this mixture and stirred for 2 h. The solvents were
removed and filtered to
137
CA 3030689 2019-01-18
yield white solid (R)-2-(4-((tert-butoxycarbonylamino)methyl)pheny1)-4,5-
dihydrothiazole-4-
carboxylic acid (38.4 g, 76.8%). Compound 67a was obtained from this acid
following the same
method as used for 38d. Yield: 84.4 %.1H NMR (500 MHz, CDC13) 87.75 - 7.77 (d,
2 H, J = 7.5
Hz), 7.27 - 7.26 (d, 2 H, J 7.5 Hz), 7.23 (s, I H), 5.62 (br, 1 H), 4.87 (br,
1 H), 4.30 (br, 2 H),
3.86 (s, 3 H), 3.78 (t, J = 10.0 Hz, 1 H), 3.48 - 3.4 (in, 1 H), 3.25 (s, 3
H), 1.42 (s, 9 H). MS (ES!)
= m/z 402.1(M + Na)t, 378.0 (M - H).
[00320] tert-Butyl 4-(4-(3,4,5-trimethoxybenzoypthiazol-2-
yl)benzylcarbamate (68a).
A mixture of 67a (2.5 mmol), CBrCI3 (3.2 mmol) and DBU (5.0 mmol) in CH2C12
(20 mL) was
stirred overnight. The reaction mixture was absorbed on silica gel and
purified by column
chromatography to yield an intermediate thiazole Weinreb amide. To a solution
of (3,4,5-
trimethoxyphenyl)magnesium bromide (0.5 M, 5.5 mL) in TI-IF was added a
solution of the
intermediate thiazole Weinreb amide (1.83 mmol) in 10 triL THF under 0 C and
stirred for 30
min. The reaction mixture was quenched with satd. NH4CI, extracted with ethyl
ether, dried with
MgSO4. The solvent was removed under reduced pressure to yield a crude
product, which was
purified by column chromatography to obtain pure compound as a light yellow
solid (32.3 %). 11-1
NMR (300M, CDCI3) 68.27 (s, I H), 7.98 (d, 2 H, J = 8.1 Hz), 7.78 (s, 2 H),
7.39 (d, 2 H, J = 8.1
Hz), 7.27 - 7.26 (d, 2 H, J = 7.5 Hz), 7.23 (s, 1 H), 4.93 (br, I H), 4.37
(br, d, 1 H), 3.96 (s, 3 H),
3.95 (s, 6 H), 1.47 (s, 9 H); MS (ES!) m/z 507.1(M + NO+.
[00321] (2-(4-(Aminomethyl)phenyl)thiazol-4-y1)(3,4,5-
trimethoxyphenyl)methanone
hydrochloride (2r or 2r-HCI). At 0 C, to a solution of 68a (200 mg) in 10 mL
CH2C12 was
added a solution of HCl in 1,4-dioxane (4 N, 2 mL) and stirred at RT for 4 h.
The precipitate (2r)
was filtered and washed with diethyl ether. Yield: 81.3%. Ili NMR (500 MHz,
DMSO-d6) 68.68
(s, 1 H), 8.38 (br, 3 H), 8.10 (d, 2 H, J = 8.4 Hz), 7.66 (d, 2 H, J = 8.4
Hz), 7.62 (s, 2 H), 4.11 (s,
2 H), 3.87 (s, 6 1-1), 3.80 (s, 3 H). MS (ES!) /ilk 385.1 (M + H)+.
(2-(44(Dimethylamino)methyl)phenyl)thiazol-4-y1)(3,4,5-
trimethoxyphenyl)methanone
hydrochloride (2u or 2u-HCl) [Figure 5]
138
CA 3030689 2019-01-18
OMe
0
OMe
S N OMe
NMe2 HCI (2u-HCI)
[00322] tert-Butyl
methyl(4-(4-(3,4,5-trimethoxybenzoyl)thiazol-2-
yl)benzyl)carbamate (71a). At 0 C, to a solution of compound 68a (100 mg, 0.2
mmol) in 5 mL
DMF was added sodium hydride (10 mg, 0.2 mmol), then iodomethane (77 mg, 0.4
mmol) was
added to the reaction mixture and stirred at RT overnight. The mixture was
quenched with a sat.
NaHCO3 solution, extracted with ethyl acetate and dried with MgSO4. The
solvent was removed
under reduced pressure to yield a crude product, which was purified by column
chromatography
to obtain pure compound 71a. Yield:.61.3%. NMR
(500 MHz, DMSO-d6) 8 8.30 (s, 1 H),
8.02 (d, 2 H, .1 = 8.0 Hz), 7.82 (s, 2 H), 7.36 (br, 2 H), 4.50 (s, 2 H), 4.00
(s, 3 H), 3.98 (s, 6 H),
2.90 (d, br, 3 H), 1.50 (s, 9 H). MS (ESI) m/z 521.2 (M + Na), 496.9 (M -
[00323] (2-(4-((Methylamino)methyl)phenypthiazol-4-y1)(3,4,5-
trimethoxyphenyl)methanone hydrochloride (2s or 2s-HCI). At 0 C, to a
solution of 71a (60
mg) in 5 mL CH2C12 was added a solution of HO in 1,4-dioxane (4 N, 2 mL) and
stirred at RT
for overnight. The precipitate (2s-HCI) was filtered and washed with diethyl
ether. Yield: 81.3%.
11-1 NIMR (500 MHz, CD03) 8 10.0 (s, 1 H), 8.29 (s, 1 H), 8.05 (d, 2 H, J =
6.0 Hz), 7.74 (s, 2 H),
7.72 (d, 2 H, J = 6.0 Hz), 4.15 (s, 2 H), 3.99 (s, 3 H), 3.96 (s, 6 H), 2.61
(s, 3 H). MS (ESI) rri/z
399.1 (M + H)+,
[00324] (2-(44(Dimethylamino)methyl)phenyl)thiazol-4-y1)(3,4,5-
trimethoxyphenyl)methanone hydrochloride (2u or 2u-HCI). To a solution of 2r
(53 mg, 0.14
mmol) in 5 mL CH2C12 was added formaldehyde solution (37% in H20, 340 mg, 4.2
mmol), and
sodium cyanoborohydride (34 mg, 0.55 mmol), the reaction mixture was absorbed
on silica gel
and free base was purified after flash column (41 mg, 70.9%). At 0 C, to a
solution of free base
(41 mg) in 5 mL CH2C12 was added a solution of HCI in 1, 4-dioxane (4 N, 2 mL)
and stirred at
139
CA 3030689 2019-01-18
RT for overnight. The precipitate (2u) was filtered and washed with diethyl
ether. Yield: 71.3%.
111 NMR (500 MHz, CDCI3) 8 13.0 (s, 11-1), 8.34 (s, I H), 8.13 (d, 2 H,) = 7.0
Hz), 7.82 (d, 2 H,
J = 7.5 Hz), 7.75 (s, 2 H), 4.24 (s, 2 H), 3.99 (s, 3 H), 3.97 (s, 6 H), 2.83
(s, 6 H). MS (ESI) nilz
413.1 (M + H)+.
[00325] 2-(4-(4-(3,4,S-Trimethoxybenzoypthiazol-2-yl)phenypacetonitrile
(2n)
0 OMe
¨ 110 OMe
N
OMe
CN (2n)
[00326] 2-(4-(4-(3,4,5-Trimethoxybenzoypthiazol-2-
yl)phenyl)acetonitrile (2n) was
prepared using the same method as used of compound lh from terephthalonitrile
and cysteine.
NMR (500MHz, CDC13) 8 8.30 (s, 1 H), 8.04 (d, 2 H), 7.76 (s, 2 H), 7.46 (d, 2
H), 3.97 (s, 3 H),
3.95 (s, 611), 3.83 (s, 2 H).
[00327] Synthesis of ' (2-(4-
(Dimethylamino)phenyl)thiazol-4-y1)(3,4,5-
trimethoxyphenyl)methanone (2o)
0 OMe
OMe
S N
OMe
=
N
n3L:- 1/4CH3
[00328] (2-(4-(Dime thyl amino)phenyl)thiazol-4-y1)(3,4,5-
trimethoxYphenyl)methanone (20) was prepared using the same method as used of
compound
113 from 4-(dimethylamino)benzonitrile and cysteine. 1H NMR (300MHz, CDC13)
58.12 (s, 1 H),
7.88 (d, 2 H), 7.80 (s, 2 H), 6.73 (d, 2 H), 3.96 (s, 3 H), 3.95 (s, 6 H),
3.05 (s, 6 H); MS (ESE)
in/z 421.1(M + NO+.
140
CA 3030689 2019-01-18
Ind ly1 A ring:
Synthesis of (2-(1H-indo1-2-yl)thiazol-4-y1)(3,4,5-trimethoxyphenyl)methanone
(62a)
[Figure 51
0 OMe
OMe
S N
OMe
NH
(62a)
[00329] 1H-Indole-2-carbonitrile (60a). To a cooled solution of indole-
2-carboxylic acid
(2.0 g, 12.4 mmol) in 60 mL of anhydrous Et20 was added 1.9 mL of SOC12 (26
mmol). After
stirring for 40 min at RT, the ether was removed under reduced pressure at a
temperature not
exceeding 35 C. The obtained acyl chloride was dissolved in 40 mL of
anhydrous Et20 and the
resulting solution was added immediately to a stirred solution of liquid
ammonia in 80 ml of
Et20. The reaction mixture was stirred at RT for 24 h. The solvent was then
evaporated under
reduced pressure, and the white indole-2-carboxamide was crystallized from 50%
aq Et0H and
dried in air, after which it was dissolved in POC13 and heated under reflux
for 5 min. The cooled
solution was poured onto crushed ice and aq NH4OH was added to maintain a
basic pH. The
aqueous mixture was extracted with Et20, the extracts were dried over Na2SO4
and evaporated.
The brown indole-2-carbonitrile 60a (63.3% overall yield from indole-2-
carboxylic acid) was
obtained. '111NMR (500 MHz, CDC13) 5 8.56 (br, s, 1 H), 7.68 (d, 1 H, J = 8.0
Hz), 7.43-7.34 (m,
2 H), 7.24-7.21 (in, 2 H). MS (ESI) silk 144.0 (M + H)+, 140.8 (M -11)".
[00330] (R)-2-(1H-indo1-2-y1)-N-methoxy-N-methy1-4,5-dihydrothiazole-4-
carboxamide (61a) was synthesized using the same method as used of 38d. 67.1%
yield.
NMR (300 MHz, CDC13) 5 9.06 (s, br, 1 H), 7.64 (d, 2 H, J 8.1 Hz), 7.36-7.24
(m, 2 H), 7.12
(cit. 1 H, J = 8.1 Hz, 1.2 Hz), 6.95 (d, 1 H, J = 1.8 Hz), 5.60 (t, br, I H, J
= 8.7 Hz), 3.86 (s, 3 H),
141
CA 3030689 2019-01-18
3.78 (t, 1 H, J = 10.2 Hi), 3.58 (dd, 1 H, J = 9.0 Hz, 10.2 Hz), 3.30 (s, 3
H). MS (ESI) m/z 312.1
(M + Na), 287.9 (M
[00331] (2-(1H-indo1-2-yl)thiazol-4-y1)(3,4,5-trimethoxyphenypmethanone
(62a) was
synthesized from 61a using the same method as used for 35a. 45.8% yield. ill
NMR (500
MHz, DMSO-d6) 6 9.26 (s, 1 H), 8.11 (s, 1 H), 7.66 (d, 1 H, J = 8.0 Hz), 7.46
(s, 2 H), 7.42 (d, 1
H, J = 8.0 Hz), 7.29 (2, 1 H, J = 7.5 Hz), 7.16 (t, 1 H, J = 7.5 Hz), 7.10 (s,
1 H), 3.97 (s, 3 H),
3.93 (s, 6 H). MS (ESI) m/z 417.1 (M + Na), 392.9 (M
Synthesis of (2-(1H-indo1-5-ypthiazol-4-y1)(3,4,5-trimethoxyphenyl)methanone
(66a)
[Figure 5]
=
0 OMe
/iIII0Me
S N
OMe
HN (66a)
[00332] (R)-2-(1-(Phenylsulfony1)-11-l-indol-5-y1)-4,5-dihydrothiazole-
4-carboxylic
acid (64a). (R)-2-(1H-indo1-5-y1)-4,5-dihydrothiazole-4-carboxylic acid 63a
was synthesized
using the same method as used for 42a from 1H-indole-5-carbonitrile and used
without further
purification. To a vigorously stirring solution of 63a (1 mmol) and
tetrabutylarnrnonium
hydrogen sulfate (0.15 mmol) in toluene (10 mL) at 0 C was added 50% aqueous
sodium
hydroxide (10 mL) and sulfonyl chloride (2 mmol). The resultant solution was
stirred at RT for 6
h. Then 1 N HC1 was added to acidify the mixture to 01=2 and extracted with
CH2C12, the
organic layer was separated and dried (MgSO4); then evaporated to dryness to
yield 64a, which
.. were used in subsequent steps without further purification.
[003331 (R)-N-methoxy-N-methy1-2-(1-(phenylsulfony1)-1H-indol-5-y1)-4,5-
dihydrothiazole-4-carboxamide (65a) was prepared from 64a with the same method
as used
for 38d. 57.1% yield. 11-1 NMR (500 MHz, CDCI3) 8 7.92 (m, 2 H), 7.77 (m, 3
H), 7.51 (d, 1 H, J
142
CA 3030689 2019-01-18
3.0 Hz), 7.46 (t, 1 H), 7.35 (t, 1H), 6.61 (d, 1 H), 5.58 (br, t, I H) 3.82
(s, 3 H), 3.73 (t, 1 H),
3.43 (m, 1 H), 3.21 (s, 3 H). MS (ESI) m/z 452.1 (M + Na)+.
[00334] (2-(1H-indo1-5-yl)thiazol-4-y1)(3,4,5-
trimethoxyphenyl)methanone (66a). To a
solution of n-BuLi (1.6 M, 1.7 mL) in 8 mL THF was added a solution of 3,4,5-
trimethoxybromobenzene (2.47 mmol) in 3 mL THF under -78 C. The mixture was
allowed to
stir for 2h and a solution of Weinreb amide 65a (1.24 mmol) in 3 mL THF was
charged. The
temperature was allowed to increase at RT and stirred overnight. The reaction
mixture was
quenched with satd. NH4C1, extracted with ethyl ether, dried with MgSO4. The
solvent was
removed under reduced pressure to yield a crude product, which was refluxed in
1 N NaOH in 5
mL ethanol solution to obtain the deprotected compound 66a and purified by
column
chromatography to obtain pure compound as a light yellow solid (36.3 %).
NMR (300M,
CDC13) 8 8.36 (br, s, 1 H), 8.31 (s, 1 H), 8.21 (s, I H), 7.92, 7.89 (dd, 1 H,
J = 1.8, 2.7 Hz), 7.46
(d, 1 H, )7.62 (s, 2 H, J ---- 8.7 Hz), 7.29 (t, 1 H, J = 2.7 Hz), 6.64 (br, 1
H), 3.97 (s, 6 H), 3.97 (s,
3 H); MS (ES1) m/z 417.1(M + Na), 392.9 (M - H)".
[00335] Synthesis of (2-(1H-Indo1-2-ypthiazol-4-y1)(1H-indol-2-y1)methanone
(8).
0
N
S N H
HN
(8)
[00336] (2-(1H-Indo1-2-yl)thiazol-4-y1)(1H-indol-2-ypmethanone (8) was
prepared
using the similar method as used of compound lh from 2-(1H-indo1-2-y1)-4,5-
dihydrothiazole-4-
carboxylic acid and cysteine. NMR
(500MHz, CDC13) 8 9.39 (s, 1 H), 8.54 (s, 1 H), 8.46 (s, 1
H), 8.06 (s, 1 H), 8.03 (dd, 1 H), 7.66 (d, 1 H), 7.51 (d, 1 H), 7.41 (d, 1
H), 7.33 (t, I H), 7.29 (d,
1 H), 7.15 (t, 1 H), 7.09 (d, 1 H), 6.72 (s, 1 H). MS (ES1) rn/z 366.1(M +
Na), 341.9 (M - H).
[00337] Synthesis of (2-(111-indo1-2-ypthiazol-4-y1)(1H-indol-5-
y1)mithanone (21).
143
CA 3030689 2019-01-18
0
NH =
S N
HN
(21)
[00338] (2-(11i-indol-2-yl)thiazol-4-y1)(1H-indol-5-yl)methanone (21)
was prepared
using the similar method as used of compound lh from 2-(1H-indo1-2-y1)-4,5-
dihydrothiazole-4-
carboxylic acid and cysteine. NMR (500MHz, CDC13) 8 9.60 (s, 1 H), 9.26 (s,
1 H), 8.31 (s, I
H), 8.03 (s, 1 H), 7.83 (dd, 1 H), 7.69 (d, 1 H), 7.53-7.49 (m, 2 H), 7.41 (t,
1 H), 7.33 (t, 1 H),
7.21-7.18 (m, 2 H), 7.13 (s, 1 H). MS (ES1) rniz 366.1(M + Na), 341.9 (M -
EXAMPLE 4
SYNTHESIS OF COMPOUNDS OF THIS INVENTION HAVING A NITROGEN
LINKER (X=NH)
[003391 To improve bioavailability, an NH linker was introduced between A
phenyl and B
thiazole rings. This new series of compounds was synthesized as shown in
Figure 6. Reaction of
3-bromo-2-oxopropanoic acid ethyl ester and arylthiourea in ethanol under 65
C produced 2-
- (arylamino)-thiazole-4-carboxylic acids 73a-d with high yields. These acids
were converted to
Weinreb amides 74a-d, followed by reactions with 3,4,5-trimethoxphenyllithium
that yielded
aniline linked free bases 5a-d, which can be converted into HO salts 5Ha-d.
Synthesis of (2-(Phenylamino)thiazol-4-y1)(3,4,5-trimethoxyphenyl)methanone
derivatives
(5a-d) and their HC1 salt [Figure 6]
OMe 0¨
OMe OMe 0 0
0 0
OMe OMe OMe 0
sN OMe N OMe SN OMe SN O¨
A
HN io HN HN = HN
=Me QV CI
(5a) (5b) (Sc) (5d)
144
CA 3030689 2019-01-18
OMe
OMe
OMe 0
0
0 OMe
OMe
Ome
OMe
N OMe s
OMe
S
HCI-HN HCI-HN
(5Ha) (5Hb) (5Hc)
[00340] General procedure for the synthesis of 2-(arylarnino) thiazole-
4-carboxylic
acids (37a-d). N-Aryl thiourea (0.01 mol) and ethyl bromopyruvate (0.011 mol)
were dissolved
in 3 mL ethanol and held at reflux for 2 h. The reaction was cooled, the
crystalline ethyl 2-
(substituted phenylamino) thiazole-4-carboxylate were collected by filtration
and washed with
ethanol. Refluxing the mixture of ethyl esters with the NaOH-ethanol solution
gave final
compounds 73a-d which were used directly in the next steps.
[00341] N-Methoxy-N-methyl-2-(arylarnino)thiazole-4-carboxamides (74a-
d) were
synthesized using the same method as used for 38d (see Example 1, Figure 2).
[00342] N-Methoxy-N-methy1-2-(phenylarnino)thiazole-4-carboxamide
(74a). 90.2%
yield. 11-1 NMR (500 MHz, CDC13) 6 7.39 (s, 2 H), 7.38 (br, 1 H), 7.36-7.33
(m, br, 4 H), 7.09 (t,
br, 1 H), 3.77 (s, 3 H), 3.43 (s, 3 H), 2.33 (s, 3 H). MS (ES1) m/z 286.0 (M +
Na)+.
[00343] N-Methoxy-N-methyl-2-(p-tolylamino)thiazole-4-carboxamide
(74b). 93.3%
yield. 11-1 NMR (500 MHz, CDC13) 8 7.35 (s, 1 H), 7.31 (br, 1 H), 7.22 (d, 2
H), 7.16 (d, 2 H),
3.76 (s, 3 H), 3.42 (s, 3 H), 2.33 (s, 3 H). MS (EST) m/z 278.0 (M + H)4.
[00344] 2-(4-Fluorophenylamino)-N-methoxy-N-methyl thiazole-4-
carboxamide (74c),
89.7% yield. 1H NMR (500 MHz, CDCI3) 37.36 (s, 1 H), 7.36-7.31 (m, 2 H), 7.07-
7.04 (m, 6 H),
3.76 (s, 3 H), 3.42 (s, 3 H). MS (ESI) m/z 282.0 (M + Na)', 280.8 (M - H).
[00345] 2-(4-Chlorophenylamino)-N-methoxy-N-methylthiazole-4-carboxamide
(74d).
'H NMR (500 MHz, CDC13) 67.66 (s, br, 1 H), 7.41 (s, 1 H), 7.34 (d, 2 H), 7.29
(d, 2 H), 3.76 (s,
3 H), 3.42 (s, 3 H). MS: 295.8 (MA); 320.0 (M+Na).
=
[00346] General procedure for the synthesis of (2-(arylamino)thiazol-4-
y1)(3,4,5-
trimethoxyphenyl)methanones (5a-d). At -78 C, to a solution of 5-bromo-1,2,3-
trimethoxybenzene (1.235 g, 5.0 mmol) in 30 mL THF was charged n-BuLi in
hexane (2.5 N, 2.4
145
CA 3030689 2019-01-18
mL, 6 mmol) under Ar2 protection and stirred for 10 min. Weinreb amide 74a-d
(1 mmol) in 10
mL THF was added to the lithium reagent and allowed to stir at RT for 2 hs.
The reaction mixture
was quenched with satd. NH4C1, extracted with ethyl ether, dried with MgSO4.
The solvent was
removed under reduced pressure to yield a crude product, which was purified by
column
chromatography to obtain pure compound (5a-d).
[003471 (2-(Phenylamino)thiazol-4-y1)(3,4,5-trimethoxyphenypmethanone
(5a). 33.3%
yield. ill NMR (500 MHz, DMSO-d6) 8 10.4 (s, 1 H), 7.85 (s, 1 H), 7.68 (d, 2
H, J = 8.0 Hz),
7.31 (t, 2 H, J = 8.0 Hz), 6.98 (t, 1 H, J = 8.0 Hz), 3.83 (s, 6 H), 3.78 (s,
3 H). MS (ESI) m/z
393.1 (M + H)+, 368.9 (M
[00348] (2(p-Tolylamino)thiazol-4-y1)(3,4,5-trimethoxyphenypinethanone
(5b). 40.6%
- yield. ill NMR (500 MHz, CDC13) 8 7.48 (s, 1 H), 7.47 (s, 2 H), 7.30 (br, 1
H), 7.27 (d, 2 H, J =-
8.5 HZ), 7.17 (d, 2 H, J 8.5 Hz), 3.93 (s, 3 H). 3.90 (s, 6 H), 2.34 (s, 3 H).
MS (ESI) m/z 385.1
(M + H)+, 382.9 (M
[00349] (2-(p-Fluorophenylarnino)thiazol-4-y1)(3,41,5-
trimethoxyphenyHmethanone
(Sc). 39.6% yield. II-1 NMR (500 MHz, CDC13) 67.52 (br, 1 H), 7.49 (s, 1 H),
7.45 (s, 2 H), 7.40-
7.37 (q, 2 H, J = 4.5 Hz), 7.08-7.04 (t, 2 H, J = 8.0 Hz), 3.93 (s, 3 El),
3.89 (s, 6H). MS (ESI) m/z
389.3 (M + H)+, 386.9 (M
[00350] (24(4-Chlorophenypamino)thiazol-4-y1)(3,4,5-
trimethoxyphenyl)methanone
(5d) was prepared using the same method as used for 5a from 1-(4-
chlorophenyl)thiourea and
ethyl bromopyruvate. Melting point: 165-166 C. 11-1 NMR (500 MHz, CDC13) 67.60
(s, br, 1 H),
7.56 (s, 1 H), 7.47 (s, 2 H), 7.38 (d, 2 H), 7.31 (d, 2 H), 3.94 (s, 3 H),
3.89 (s, 6-H). MS: 402.9
(M-l); 427.0 (M+Na)t
[00351] General procedure for the synthesis of hydrochloride salts (5Ha-
c). At 0 C, to
a solution of compound 5a-c (0.1 mmol) in 5 mL CH2Cl2 was added a solution of
HC1 in 1,4-
dioxane (4 N, 2 mL) and stirred at RT for overnight. The precipitates 5Ha-c
were collected and
washed with diethyl ether.
[00352] (2-(Phenylarnino)thiazol-4-y1)(3,4,5-trimethoxyphenyl)methanone
hydrochloride salt (5Ha). 91.6% yield. II-I NMR (500 MHz, DMSO-d6) 8 12.9 (br,
1 H), 7.49-
7.46 (m, 2 H), 7.42-7.40 (m, 2 H),7.37-7.34 (m, br, 2 H), 7.11 (s, 2 H), 3.94
(s, 3 H), 3.92 (s, 6
H). MS (ESI) m/z 389.1 (M + H)+.
146
CA 3030689 2019-01-18
[00353] (24p-Tolylamino)thiazol-4-y1)(3,4,5-trimethoxyphenyl)methanone
hydrochloride salt (5Hb). 39.6% yield. 1H NMR (500 MHz, CDCI3) 8 7.30-7.25 (m,
br, 5 H),
7.12 (s, 2 H), 3.94 (s, 3 H), 3.92 (s, 6 H), 2.38 (s, 3 H). MS (ES!) m/z 389.1
(M + H)*.
[00354] (24p-Fluorophenylamino)thiazol-4-y1)(3,4,5-
trimethoxyphenyl)methanone
hydrochloride salt (5Hc). 89.3% yield. 1H NMR (500 MHz, CDC13) 8 10.55 (s, 1
H), 7.85 (s, 1
H), 7.72-7.69 (q, 2 H, J = 4.5 Hz), 7.50 (s, 2 H), 7.18-7.15 (t,2 H, J = 8.5
Hz), 4.30 (br, 1 H),
3.82 (s, 611), 3.78 (s, 3 H). MS (ES!) m/z 389.3 (M + H)+.
Synthesis of (2-(Phenylamino)-1H-intidazol-4-y1)(3,4,5-
trimethoxyphenyl)methanone (5e)
o
Lo CH3S0
NH
31-1 ( cr
Et0H _____________________________________________ HN N
BrCN
. H,
C 38% HN--C--t4 HN
b 1111
a
HOt
H N mCI
HN
0¨ 0-
0 0
t-BuLl ¨ HC1 0
0¨
o¨ H N 0¨
Ni"
Et20 HN 140
¨"o-
eSe
Scheme 1: preparation of compound 5e.
[00355] 2,2-Diethoxy-N-(iminomethylene)ethanamine (a)., A solution of
the
aminoacetaldehyde diethyl acetal (5.32 g, 40 mmol) in ether (20 mL) was added
to a suspension
of CNBr (4.22 g, 40 mmol) in hexane (20 mL) at RT. The reaction mixture was
stirred at RT
overnight. The solid was removed by filtration and washed with ether. The
combined filtrate was
concentrated. Flash chromatography of the concentrated residue afforded 2.82 g
(45%) of the N-
(2,2-diethoxyethyl)carbodiimide (a). 1H NMR (500 MHz, CDCI3): 4.58 (t, J 5.5
Hz, 1 H), 3.85
147
CA 3030689 2019-01-18
(br s, 1 H), 3.73 (m, 2 H), 3.56 (m, 2 H), 3.16 (J = 5.5 Hz, 2 H), 1.23 (1,1
7.0 7.0 Hz, 3 H), MS:
156.8 (M-H); 180.9 (M+Na).
[00356] 1-(2,2-Diethoxyethyl)-3-phenylguanidine (b). Aniline (1.66 g,
17.8 mmol) was
dissolved in ethanol (25 mL), and N-(2,2-diethoxyethyl)carbodiimide (a), (2.82
g, 17.8 mmol),
was added dropwise. Then methanesulfonic acid (1.71 g, 17.8 mmol) was added,
and the mixture
was warmed at reflux for 24 h. The reaction mixture was poured into NaOH (0.5
M) and
extracted with CH2C12. Drying and concentration afforded a product that was
subjected to flash
chromatography to give the intermediate guanidine (b) (3.3 g, 73.8%). 'H NMR
(500 MHz,
DMSO-d6) 8 7.27-6.90 (m, 5 H), 4:55 (t, 1 H), 3.76-3.70 (m, 2 H), 3.60-3.54
(m, 2 H), 3.35-3.34
.. (d, 2 H), 1.22 (pent, 6 H). MS: 249.8 (M-H); 252.1(M+H)+.
[00357] N-Pheny1-1H-imidazol-2-amine (c). The guanidine (b) was
dissolved in HC1 (5
mL, 6 M) at 0 C and then stirred for 2 h. After the starting material was
consumed, NaOH (25%)
was added until a precipitate formed. This mixture was stirred for 30 min. The
reaction was then
poured into NaOH (0.5 M), extracted with CH2C12, dried and concentrated. Flash
chromatography afforded (c) (0.95 g, 50 %). 'H NMR (500 MHz, DMSO-d6) 8 8.58
(s, br, 1 H),
7.34-6.74 (m, 5 H), 6.68 (s, 2 H), 6.62 (br, 2 H), 3.82 (s, 6 H), 3.73 (s, 3
H). MS: 157.6 (M-H);
160.0 (M+H) .
[00358] N-Phenyl-1-trity1-1H-imidazol-2-amine (d). Trityl chloride
(2.79 g, 10 !mop
was added to an ice-cooled solution of phenyl amino imidazole (c) ( 1.59 g, 10
mmol) and
triethylamine (1.01 g, 10 mmol) in methylene dichloride (50 mL). The reaction
'mixture was
allowed to warm to RT and stirred overnight. The mixture was diluted with
methylene dichloride,
washed successively with H20, saturated NaHCO3, brine and dried with MgSO4.
Filtration and
evaporation of the solvent followed by chromatography separation gave the
product (d). NMR
(500 MHz, CDC13) 67.52-7.35 (m, 5 H), 7.28-7.43 (m, 15 H), 6.85 (s, 2 H), 6.41
(s, 1 H), 6.08 (s,
1 H). MS: 1399.8 (M-H); 402.8 (M+H)+.
[00359) (2-(Phenylamino)-1-trity1-1H-imidazol-4-y1)(3,4,5
trimethoxyphenyl)methanone (e). At -78 C, t-BuLi in THF (1.7 M, 0.34 mL, 0.58
mmol) was
added to a solution of trityl protected compound (d) (116 mg, 0.289 mmol) in
THF. Then 3,4,5-
trimethoxybenzoyl chloride (66.5 mg, 0.289 mmol) was added and stirred
overnight. The reaction
mixture was quenched with saturated NH4CI, and dried with MgSO4. Filtration
and evaporation
of the solvent followed by chromatography afforded compound (e) (75 mg,
43.7%).11-1 NMR
148
CA 3030689 2019-01-18
(500 MHz, CDC13) 8 7.55-7.41 (m, 5 H), 7.32 (s, 1 H), 7.28-7.18 (m, 15 H),
6.94 (s, 2 H), 3.78 (s,
6 H), 3.70 (s, 3 H). MS: 594.2 (M-H); 596.3 (M+H).
[00360] (2-
(Phenylamino)-1H-imidazol-4-y1)(3,4,5-trimethoxyphenyl)methanone (5e).
To a solution of trityl protected compound (e) (50 mg, 0.084 mmol) in ethyl
ether was added 2 M
HCI in ether (1 mL, 1 mmol). The reaction mixture was stirred overnight and
washed with
saturated NaHCO3 and dried with MgSO4. Filtration and evaporation of the
solvent followed by
flash chromatography to yield de-protection compound 5e (18 mg, 63%). NMR
(500 MHz,
DMSO-d) 57.54 (s, br, 1 H), 7.51-743 (m, 3 F1), 7.33 (d, 2 H), 7.04 (s, 2 H),
6.62 (br, 2 H) 3.82
(s, 6 H), 3.73 (s, 3 I-1). MS: 352.1 (M-H)-; 354.3 (M+H).
EXAMPLE 5
SYNTHESIS OF SELECTED ARYL-BENZO'YL-IMIDAZOLE COMPOUNDS
IS
0
R1(1 )
________________________________________________ HN 2
Preparation of 2-aryl-4,5-dihydro-1H-imidazoles 14b, 14c, 14x (Figure 7).
R, ________________________________
HN
[00361] To a
solution of appropriate benzaldehyde 8(b, c, x) (60 mmol) in t-BuOH (300
mL) was added ethylenediamine (66 mmol) and stirred for 30 min at RT.
Potassium carbonate
(75 mmol) and iodine (180 mmol) were added to the reaction mixture
sequentially followed by
stirring at 70 C for 3 h. Sodium sulfite (Na2S03) was added and the mixture
was extracted by
chloroform. The organic layer was dried over magnesium sulfate and
concentrated. The residue
was purified by flash column chromatography (chloroform: methanol 20:1) to
give a white solid.
Yield: 50-60%.
149
CA 3030689 2019-01-18
Preparation of 2-aryl-1H-imidazoles (9a-j, p, x; Figures 7 and 8).
I
[00362] Method A (essential for only 9b, 9x Figure 7): To a solution of
2-ary1-4,5-
dihydro-1H-imidazole 14b, x (35 mmol) in DMSO (100 mL) was added potassium
carbonate
(38.5 mmol) and diacetoxyiodobenzene (38.5 mmol). The reaction mixture was
stirred overnight
in darkness. Water was added followed by extraction with dichloromethane. The
organic layer
was dried over magnesium sulfate and concentrated. The residue was subjected
to flash column
chromatography (hexane: ethyl acetate 3:2) to give a white solid. Yield: 30%-
50%.
[00363] Method B (essential for only 9c; Figure 7): To a solution of 2-ary1-
4,5-dihydro-
1H-imidazole 14c (50 mmol) in DMF (70 mL) was added DBU (55 mmol) and CBrC13
(55
mmol). The reaction mixture was stirred overnight and a saturated NaHCO3
(aqueous) solution
was added followed by extraction with dichloromethane. The organic layer was
dried over
magnesium sulfate and concentrated. The residue was subjected to flash column
chromatography
(chloroform: methanol 50:1) to yield a white solid. Yield: 7%.
[00364] Method C (essential for 9a, 9d-j, 9p; Figure 8): To a solution
of appropriate
benzaldehyde (8a, 8d-j, 8p) (100 mmol) in ethanol (350 mL) at 0 C was added a
solution of
40% oxalaldehyde in water (12.8 mL, 110 mmol) and a solution of 29% ammonium
hydroxide in
water (1000 mmol, 140 mL). After stirring for 2-3 days at RT, the reaction
mixture was
concentrated and the residue was subjected to flash column chromatography with
dichloromethane as eluent to yield the titled compound as a yellow powder.
Yield: 20%- 40%.
Preparation of 2-aryl-14phenyisulfony1)-1H-imidazoles (10a-j, p, x; Figures 7
and 8).
j
Ph
[00365] To a solution of 2-aryl-1H-imidazole 9a-j, p, x (20 mmol) in
anhydrous THF (200
mL) at 0 'V was added sodium hydride (60% dispersion in mineral oil, 1.2 g, 30
mmol) and
stirred for 30 min. Benzenesulfonyl chloride (2.82 mL, 22 mmol) was added and
the reaction
150
CA 3030689 2019-01-18
mixture was stirred overnight. After dilution by 100 mL of saturated NaHCO3
solution (aqueous),
the reaction mixture was extracted by ethyl acetate (500 mL). The organic
layer was dried over
magnesium sulfate and concentrated. The residue was purified by flash column
chromatography
(hexane: ethyl acetate 2:1) to give a pale solid. Yield: 50%-70%.
Preparation of aryl (2-aryl-1-(phenylsulfonyl)-1H-imidazol-4-yl)methanones (I
laa-ai, ba, ca,
cb, da, db, ea, eb, fa, fb, ga, gb, ha, hb, to, ib, ja, jb, pa; Figures 7 and
8).
0
Ph
[00366] To a solution of 2-aryl-1-(phenylsulfony1)-1H-imidazole (6.0 mmol)
10a-j, p, x in
anhydrous THF (30 mL) at -78 C was added 1.7M tert-butyllithium in pentane
(5.3 mL, 9.0
mmol) and stirred for 10 min. Appropriate substituted benzoyl chloride (7.2
mmol) was added at
-78 C and stirred for overnight. The reaction mixture was diluted with 100 mL
of saturated
NaHCO3 solution (aqueous) and extracted by ethyl acetate (200 mL). The organic
layer was dried -
over magnesium sulfate and concentrated. The residue was purified by flash
column
chromatography (hexane: ethyl acetate 4:1) to give a white solid. Yield: 15%-
40%.
General procedure for the preparation of aryl (2-aryl-1H-imidazol-4-
yl)methanones (12aa-ai,
ba, ca, cb, da, db, ea, eb, fa, fb, ga, gb, ha, hb, ia, ib, ja, jb, pa;
Figures 7 and 8).
0
R _________________________
2¨R
HN
[00367] To a solution of aryl (2-aryl-1-(phenylsulfony1)-1H-imidazol-4-
yl)methanones
(2.0 mmol) llaa-ai, ba, ca, cb, da, db, ea, eb, fa, fb, ga, gb, ha, hb, ia,
ib, ja, jb, pa in TI-IF (20.0
mL) was added 1.0M tetrabutyl ammonium fluoride (4.0 mmol) and stirred
overnight. The
reaction mixture was diluted by 50 mL of saturated NaHCO3 solution (aqueous)
and extracted by
ethyl acetate (100 mL). The organic layer was dried over magnesium sulfate and
concentrated.
151
CA 3030689 2019-01-18
The residue was purified by flash column chromatography (hexane: ethyl acetate
3:1) or
recrystallized from water and methanol to give a white solid. Yield: 80-95%.
Preparation of (2-(4-hydroxypheny1)-1H-imidazol-4-y1) (aryl)methanones (12ka,
12kb; Figure
8).
0
HO
I -1 112
HN
1003681 To a solution of (2-(4-(benzyloxy)pheny1)71H-imidazol-4-
y1)(aryl)methanone 12ja
or 12jb, (1 mmol) in AcOH (20 mL) was added concentrated HC1 (2 mL) and
refluxed overnight.
After removing the solvent, the residue was recrystallized from
dichloromethane to give the titled
compound as a yellow solid. Yield: 70-85%.
Preparation of (2-aryl-1H-imidazol-4-y1) (3,4,5-trihydroxyphenyl)methanones
13ea, 13fa, 13ha
(Figure 8).
0
OH
HN
OH
OH
[00369] To a solution of aryl (2-aryl-1H-imidazol-4-yl)methanone 12ea,
12fa or 12ha (0.5
mmol) in CH2C12 (6.0 mL) was added 1.0 M of BBr3 (2 mmol) in CH2Cl2 and
stirred for 1 h at
RT. Water was added to destroy excess BBr3. The precipitated solid was
filtered and
recrystallized from Me0H to afford a yellow solid. Yield: 60-80%.
Preparation of aryl (2-aryl-1H-imidazol-4-yl)methanone-HC1 salt (12db-HC1).
0
Ri _________________________
_________________________________________________ R2
HCI HN
= 152
CA 3030689 2019-01-18
[00370] To a solution of 12db (0.5 mmol) in methanol (20 mL) was added
2 M solution of
hydrogen chloride (5 mmol) in ethyl ether and stirred overnight at RT. The
reaction mixture was
concentrated and the residue was washed by CH2Cl2 to yield the titled
compound. Yield: 95%.
Preparation of aryl (2-phenyl-1H-imidazol-1-Amethanone (12aba, 12aaa; Figure
9).
R, /-
0
R2
[003711 To a solution of 2-phenyl- I H-imidazole 9a (10 mmol) in THF
(20 mL) was added
NaH (15 mmol) and substituted benzoyl chloride (12 mmol) at 0 C. The reaction
mixture was
stirred overnight and diluted by saturated NaHCO3 solution followed by
extraction with ethyl
acetate. The organic layer was dried over magnesium sulfate and concentrated.
The residue was
purified by flash column chromatography (chloroform) to give a white solid.
Yield: 12-16%.
Preparation of 1-substituted-(2-pheny1-1H-imidazol-1-y1)-aryl-methanone (12dc,
12fc, 12daa,
12 dab, 12 cba, 11gaa, 12k; Figures 10-11).
0
R __________________________
<, '\)
_________________________________________________ R2
R2
[00372] The synthesis of 12dc, 12k and 12daa, 12dab and 12eba is
summarized in
Figure 10. Compounds 12da, 12cb and 12fa were synthesized according to the
synthesis
decribed above and in Figures 7 and 8. Treatment of 12da and 12fa with
aluminum chloride
153
CA 3030689 2019-01-18
provided the para-demethylated 12dc, 12fc with the 3,5-dimethoxy being
intact.. Compound
12daa was prepared by benzylation of the N-1 position of 12da. While
methylation of the N- I
position of 12da and 12cb afforded compounds 12dab and 12cba, respectively.
[00373] Synthesis of 12dc, 12fc, 12daa, 12dab, 12cba: Method D. (for
12dc and 12fc)
[Figure 10j:
OMe
Ri
HN
OH
OMe
RI=CH3 (12dc)
RI=C1 (12fc)
[00374] To a solution of 12da and 121a (200 mg) in THF (20 mL) was added
aluminum
chloride (10 equiv). The reaction mixture was stirred overnight. Water was
added followed by
extraction with ethyl acetate. The organic layer was dried over magnesium
sulfate and
concentrated. The residue was subjected to flash column chromatography
(hexane: ethyl acetate
1:1) to give a white-yellowish solid. Yield: 60%-80%.
Synthesis of 12daa, 12dab, 12cba, Method E: [Figure 10]:
RI
IN
R2
RI=Me; R2=Bn; R3=3,4,5-(0Me)3(12daa)
R2=Me; R2=CH3; R3=3,4,5-(0Me)3(12dab)
R1=0Me; R2=CH3; R3=F (12cba)
[00375] To a solution of 12da and 12cb (100 mg) in THF (10 mL) in an
ice-bath was
added sodium hydride (1.2 equiv) followed by the addition of methyl iodide
(for 12dab, 12cba)
or benzyl bromide (for 12daa) (2 equiv). The resulted reaction mixture was
stirred for 5 h under
154
CA 3030689 2019-01-18
reflux condition. After dilution by 50 mL of saturated Na11CO3 solution.
(aqueous), the reaction
mixture was extracted by ethyl acetate (100 mL). The organic layer was dried
over magnesium
sulfate and concentrated. The residue was purified by flash column
chromatography (hexane:
ethyl acetate 2:1) to give a white solid. Yield: 50%-98%.
Synthesis of llgaa and 121a (Figure 11):
0
OMe
Ri
N
OMe
R2
OMe
RI=N(Me)2; R2=--(4-0Me)PhS02 (1 1 gaa)
RI=13r; R2=H (121a)
[00376] The substituted benzaldehyde compounds 8(1, g) were converted
to compounds
9(1, g) in the presence of ammonium hydroxide and glyoxal to construct the
imidazole scaffold.
The imidazole rings of compounds 9(1, g) were protected by an appropriate
phenylsulfonyl group
followed by coupling with 3,4,5-trimethoxybenzoyl chloride to achieve compound
11(la,gaa).
Treatment of 111a with tert-butylammoniumfluoride to remove the protecting
group afforded
121a.
[00377] Structural characterization of (1-Benzy1-2-(p-toly1)-1H-imidazol-4-
y1)(3,4,5-
trimethoxyphenyl)methanone (12daa) (Figure 11).
=0
0
--N
0
(12daa)
[00378] Yield: 92.8%; mp 135-137 C. NAV
(CDC13, 500 MHz) 8 7.81 (s, 1 H), 7.80
(d, J = 6.5 Hz, 2 H), 7.58 (d, J = 8.0 Hz, 2 H), 7.41-7.45 (m, 3 H), 7.31-7.33
(m, 2 H), 7.20 (d, J
-,-- 7.0 Hz, 2 H), 5.33 (s, 2 H), 3.99 (s, 3 H), 3.98 (s, 6 H), 2.47 (s, 3 H).
MS (ESI) calcd for
C27H26N204 442.2, found 443.1 [M + Na]. HPLC I: tR 4.28 mm, purity > 99%.
155
CA 3030689 2019-01-18
[00379] Structural characterization of (2-(4-(dirnethylamino)pheny1)-
14(4.
methoxyphenyl)sulfony1)-1H-imidazol-4-y1)(4-fluorophenyl)methanone (12gba).
0
It
so
0
N
(12gba)
[00380] Yield: 34.1%; mp
147-149 C. NMR (CDC13, 500 MHz) 8 8.07 (q, J = 8.5 Hz,
5.5 Hz, 2 H), 7.78 (d, J = 9.0 Hz, 2 H), 7.41 (d, J = 8.5 Hz, 2 H), 7.39 (s, 1
H), 7.23 (t, J = 8.5 Hz,
2 H), 6.91 (d, J= 9.0 Hz, 2 H), 6.68 (d, J = 9.0 Hz, 2 H), 3.89 (s, 3 H), 3.08
(s, 3 H). MS (ES!)
calcd for C25H22FN304S 479.1, found 502.1 [M + Na]. HPLC2: tR 18.6 min, purity
96.9%.
Synthesis of (2-(4-bromopheny1)-1H-iznidazol-4-y1)(3,4,5-
trimethoxYphenyl)methanone
(121a) (Figure 11)
0
OMe
Br
HN
OMe
OMe
[00381] Synthesis of 91, 9g: To a solution of appropriate benzaldehyde
(81, and 8g, 100
mmol) in ethanol (400 mL) at 0 C was added a solution of 40% oxalaldehyde
(glyoxal) in water
.. (1.1 equiv) and a solution of 29% ammonium hydroxide in water (10 equiv).
After stirring for 2-3
days at RT, the reaction mixture was concentrated and the residue was
subjected to flash column
chromatography with dichloromethane as eluent to yield the titled compound as
a yellow powder.
Yield: 10%- 30%.
[00382] Synthesis of 101a, 10gb: To a solution of irnidazoles (91, 9g)
(10 mmol) in
.. anhydrous THF (200 mL) at 0 C was added sodium hydride (60% dispersion in
mineral oil, 1.2
equiv) and stirred for 20 min. 4-Methoxybenzenesulfonyl chloride (for 10gb) or
benzenesulfonyl
156
CA 3030689 2019-01-18
chloride (for others)(1.2 equiv) was added and the reaction mixture was
stirred overnight. After
dilution by 200 mL of saturated NaHCO3 solution (aqueous), the reaction
mixture was extracted
by ethyl acetate (600 mL). The organic layer was dried over magnesium sulfate
and concentrated.
The residue was purified by flash column chromatography (hexane: ethyl acetate
2:1) to give a
pale solid. Yield: 40%-95%.
[00383] Synthesis of 111a, llgaa: To a solution of 2-ary1-1-
(phenylsulfony1)-1H-
imidazole (101a, 10gb) (5.0 mmol) in anhydrous THF (30 mL) at -78 C was added
1.7 M ten-
butyllithium in pentane (1.2 equiv) and stirred for 10 min. 3,4,5-
Trimethoxybenzoyl chloride (1.2
equiv) was added at -78 C and stirred overnight. The reaction mixture was
diluted with 100 mL
of saturated NaHCO3 solution (aqueous) and extracted by ethyl acetate (300
mL). The organic
layer was dried over magnesium sulfate and concentrated. The residue was
purified by flash
column chromatography (hexane: ethyl acetate 3:1) to give a white solid.
Yield: 5%-45%.
[00384] Synthesis of 121a: To a solution of aryl (2-aryl-1-
(phenylsulfony1)-1H-imidazol-
4-y1)methanone (11Ia), 2.0 nunol) in THF (25.0 mL) was added 1.0 M tetrabutyl
ammonium
fluoride (2 equiv) and stirred overnight. The reaction mixture was diluted by
60 mL of saturated
NaHCO3 solution (aqueous) and extracted by ethyl acetate (150 mL). The organic
layer was dried
over magnesium sulfate and concentrated. The residue wets purified by flash
column
chromatography (hexane: ethyl acetate 4:1) or recrystallized from water and
methanol to give a
white solid. Yield: 80-98%.
Synthesis of (4-Fluorophenyl)(2-(4-methoxypheny1)-1H-imidazol-4-y1)methanone
(I2cb)
(Figure 7).
Me0
HN
[00385) To a solution of (4-fluorophenyl)(2-(4-methoxypheny1)-1-
(phenylsulfonyI)-1H-
imidazol-4-yOmethanone (11cb, 872 mg, 2.0 mmol) in THF (20.0 mL) was added 1.0
M
tetrabutyl ammonium fluoride (4.0 mL, 4.0 mmol) and stirred overnight. The
reaction mixture
was diluted by 50 mL of saturated NaHCO3 solution (aqueous) and extracted by
ethyl acetate
157
CA 3030689 2019-01-18
(100 mL). The organic layer was dried over magnesium sulfate and concentrated.
The residue
was recrystallized from water and methanol to give a white solid. Yield: 90%;
mp 245 ¨ 247 C.
Synthesis of (2-(p-Toly1)-1H-imidazo1-4-y1)(3,4,5-trimethoxyphenyl)methanone
(12da)
(Figure 8).
0
OMe
Me
HN
OMe
OMe
[00386] To a solution of (1-(phenylsulfony1)-2-(p-toly1)-1H-imidazol-4-
y1)(3,4,5-
trimethoxyphenypmethanone (11da, 492 mg, 1.0 mmol) in THF (15.0 mL) was added
1.0 M
tetrabutyl ammonium fluoride (2.0 mL, 2.0 mmol) and stirred overnight. The
reaction mixture
was diluted by 30 mL of saturated NaHCO3 solution (aqueous) and extracted by
ethyl acetate (80
mL). The organic layer was dried over magnesium sulfate and concentrated. The
residue was
recrystallized from water and methanol to give a white solid. Yield: 88.5%.
Synthesis of (2-(4-Chloropheny1)-1H-imidazol-4-y1)(3,4,5-
trimethoxyphenyl)methanone
(12fa) (Figures 8 and 14).
0
OMe
CI
HN
OMe
OMe
[00387] 2-(4-Chloropheny1)1H-imidazole (90: To a solution of 4-
chlorobenzaldehyde
(8f) (100 mmol) in ethanol (350 mL) at 0 C was added a solution of 40%
oxalaldehyde in water
(12.8 mL, 110 mmol) and a solution of 29% ammonium hydroxide in water (1000
mmol, 140
mL). After stirring for 2-3 days at RT, the reaction mixture was concentrated
and the residue was
subjected to flash column chromatography with dichloromethane as eluent to
yield the titled
compound as a yellow powder. Yield: 19.8 %. 11-1 NMR (500 MHz, DMSO-d6) 8
13.60 (br, 1H),
158
CA 3030689 2019-01-18
7.94(d, J = 8.5 Hz, 21-1), 7.51 (d, J = 8.0 Hz, 2H), 7.27 (s, 1H), 7.03 (s,
1H). MS (ES!): calculated
for C9H7C1N2, 178.0, found 178.9 [M + Hr.
[00388] 2-(4-Chloropheny1)-1-(phenylsulfony1)-1H-imidazole (10f): To a
solution of 2-
(4-chloropheny1)-1H-imidazole (9f) (20 mmol) in anhydrous THF (200 mL) at 0 C
was added
sodium hydride (60% dispersion in mineral oil, 1.2 g, 30 mmol) and stirred for
30 min.
Benzenesulfonyl chloride (2.82 mL, 22 mmol) was added and the reaction mixture
was stirred
overnight. After dilution by 100 mL of saturated NaHCO3 solution (aqueous),
the reaction
mixture was extracted by ethyl acetate (500 mL). The organic layer was dried
over magnesium
sulfate and concentrated. The residue was purified by flash column
chromatography (hexane:
ethyl acetate 2:1) to give a pale solid. Yield: 54.9%. IF1 NMR (500 MHz,
CDC13) 5 7.65 (d, J =
2.0 Hz, 1H), 7.58 (t, J 7.5 Hz, 1H), 7.43 (d, J = 8.5 Hz, 2H), 7.38 (t, I =
8.0 Hz, 2H), 7.34-7.36
(m, 4H), 7.12 (d, J = 1.5 Hz, 1H). MS (ESL): calculated for C15li11CIN202S,
318.0, found 341.0
[M + Na].
[00389] (2-(4-Chloropheny1)-1-(phenylsulfony1)-1H-imidazol-4-y1)(3,4,5-
trimethoxypheny1)methanone (11fa): To a solution of 2-(4-chloropheny1)-1-
(phenylsulfony1)-
1H-imidazole (101) (6.0 mmol) in anhydrous THF (30 mL) at -78 C was added 1.7
M tert-
butyllithium in pentane (5.3 mL, 9.0 mmol) and stirred for 10 min. 3,4,5-
Trimethoxybenzoyl
chloride (7.2 mmol) was added at -78 C and stirred for overnight. The
reaction mixture was
diluted with 100 mL of saturated NaHCO3 solution (aqueous) and extracted by
ethyl acetate (200
mL). The organic layer was dried over magnesium sulfate and concentrated. The
residue was
purified by flash column chromatography (hexane: ethyl acetate 4:1) to give a
white solid. Yield:
36.8%; 1H NMR (500 MHz, CDC13) 5 8.05 (d, J = 7.5 Hz, 2H), 7.77 (t, J = 7.5
Hz, I H), 7.62 (t, J
= 8.0 Hz, 2H), 7.48 (s, 1H), 7.44 (d, J= 9.0 Hz, 2H), 7.39 (d, J 8.5 Hz, 2H),
7.37 (s, 2H). MS
(BSI): calculated for C251-121C1N206S, 512.1, found 513.1 [M + H].
[00390] (2-(4-Chloropheny1)-1H-imidazol-4-y1)(3,4,5-trimethoxyphenyl)methanone
(12fa): To a solution of (2-(4-chloropheny1)-1-(phenylsulfony1)-1H-imidazol-4-
y1)(3,4,5-
trimethoxyphenyl)methanone (Ma) (2.0 mmol) in THF (20.0 mL) was added 1.0 M
tetrabutyl
ammonium fluoride (4.0 mmol) and stirred overnight. The reaction mixture was
diluted by 50 mL
of saturated NaHCO3 solution (aqueous) and extracted by ethyl acetate (100
mL). The organic
layer was dried over magnesium sulfate and concentrated. The residue was
purified by flash
column chromatography (hexane: ethyl acetate 3:1) or recrystallized from water
and methanol to
159
CA 3030689 2019-01-18
give a white solid. Yield: 80-95%. Yield: 36.9%; mp 193 - 195 C. NMR
(500 MHz, CDC13)
8 10.75 (br, 1H), 7.96 (d, J = 85 Hz, 2H), 7.83 (s, 1H), 7.47 (d, J = 9.0 Hz,
2}1), 7.23 (s, 2H),
3.97 (s, 3H), 3.94 (s, 6H), 2.43 (s, 3H). MS (ESI): calculated for
C19141701\1204, 372.1, found
395.1 [M + Na], 370.9 [M - Hr. HPLC Gradient: Solvent A (water) and Solvent B
(methanol):
0-15 min 40-100%B (linear gradient), 15-25 min 100%B: tR 16.36 min, purity >
99%.
Synthesis of (2-(4-Chloropheny1)-1H-imidazol-4-y1)(4-fluorophenyl)methanone
(12fb) (Figure
8).
CI
HN
[00391] To a solution of (2-
(4-chloro phen y1)- 1-(phen yls ulfony1)-1H-imidazo1-4-y1)(4-
fluorophenyOmethanone (11fb, 440 mg, 1.0 mmol) in TI-IF (12.0 mL) was added
1.0 M tetrabutyl
ammonium fluoride (2.0 mL, 2.0 mmol) and stirred overnight. The reaction
mixture was diluted by
rriL of saturated NaHCO3 solution (aqueous) and extracted by ethyl acetate (60
mL). The organic
15 layer was dried over magnesium sulfate and concentrated. The residue was
recrystallized from water
and methanol to give a white solid. Yield: 83.7%.
Physicochemical Characterization of Aryl-Benzoyl-Imidazole Compounds and
Intermediates
Compound Physicochemical Cheracterization
2-phenyl-1H-imidazole (9a) - Yield: 36.8 %. 1H NMR (500 MHz, DMSO-
d6) 5 12.52 (br, 1 H), 7.95 (d, J = 7.0 Hz, 2 H),
7.44 (t, J = 7.5 Hz, 2 H), 7.34 (t, J = 7.0 Hz,
1H), 7.25-7.27m, 1 H), 7,04 - 7.07m, 1 H).
MS (ESI): calculated for C9H8N2, 144.1, found
167.1 [M + Na]+.
2-(4-fluoropheny1)-1H-i midazole (9b) Yield: 56.5 %. 'H NMR (300 MHz, DMSO-
d6) 6 12.46 (br, 1 H), 7.94-7.99 (m, 2 H), 7.24-
7.30 (m, 2 H), 7.00- 7.03 (m, 2 H). MS (ES1):
calculated for C9H7FN2, 162.1, found 163 [M
+H], 160.6 [M - HT.
2-(4-methoxypheny1)-1H-imidazole (9c) Yield: 22.2 %. 111 NMR (500 MHz,
CDC13) 5
7.80 (d, J = 10.0 Hz, 2 H), 7.15 (s, 2 H), 3.86
(s, 3 H). MS (ES!): calculated for C101-110N20,
174.1, found 175 TM + Hr, 172.8 [M - Hr.
160
=
CA 3030689 2019-01-18
2-(p-toly1)-1H-imidazole (9d) Yield: 36.1 %. 111 NMR (500 MHz, CDC13) 5
7.64 (d, J = 7.5 Hz, 2 H), 7.16(d, J= 7.5 Hz, 2
H), 7.12 (s, 1 H), 7.02 (s, 1 H). MS (ESD:
calculated for Ci0K0N2, 158.1, found 159.0
(1µ4 + Hr, 156.8 [M -- Hr.
2-(3,4,5-trimethoxypheny1)-1H-imidazole (9e) Yield: 26.0%. 11-1 NMR (500
MHz, CDC13) 8
7.26 (s, 2 H), 7.08 (d, J= 1.5 Hz, 2 H), 3.86 (s,
3 H), 3.82 (s, 6 11). MS (ESD: calculated for
Cl2H14N203, 234.1, found 234.9 (M + Hr.
=
2-(4-chloropheny1)-1H-imidazole (90 Yield: 19.8 %. 'H NMR (500 MHz, DMSO-
d6) 8 13.60 (br, 1 H), 7.94 (d, 1= 8.5 Hz, 2 H),
7.51 (d, .1= 8.0 Hz, 2 H),7.27 (s, 1 1-1), 7.03 (s.
1 H). MS (ES!): calculated for C91-17C1N2,
178.0, found 178.9 IM + Hr.
4-(1H-imidaw1-2-y1)-N,N-dimethylaniline (9g) Yield: 16,5 %. 1H NMR (300 MHz,
CDC13)
7.70 (dd, J= 7.0 Hz, 2.0 Hz, 2 H), 7.10 (s, 2
H), 6.75 (dd, J= 9.0 Hz, 2.0 Hz, 2 H), 3.02 (s,
6 H). MS (ES!): calculated for C111113N3,
187.1, found 187.9 EM + Hr, 185.8 (1µ.4 - Hr.
2-(3,4-dimethoxypheny1)-1H-imidazole (9h) Yield: 22,0 %. 'H NMR (500 MHz,
CDC13) 8
7.52 (d, J= 1.5 Hz, 1 H), 7.27-7.28 (m, 1 I-1),
7.14 (s, 2 H), 6.88 (d, J= 8.0 Hz, 1 H), 3.91 (s,
3 H), 3.87 (s, 3 H). MS (ESL): calculated for
C111-112N202, 204.1, found 205.1 [M + Hr, =
202.8 [M - Hr.
2-(2-(trifluoromethyl)pheny1)-1H-imidazole Yield: 25.5 %. NMR (500 MHz,
DMS045)
(90 8 12.31 (br, 1 H), 7.84 (d, J= 8.0 Hz, 1
H), 7.76
J= 8.0 Hz, 1 H), 7.65 (t, J = 7.5 Hz, 1 H),
7.16 (br, 2 H). MS (al): calculated for
Ci0H7F3N2, 212.1, found 212.9 [M + lir, 210.7
- HT.
2-(4-(benzyloxy)pheny1)-1H-imidazole (9j) Yield: 12.1 %. 'H NMR (500 MHz,
CDC13)
7.77 (d, J= 8.5 Hz, 2 H), 7.36-7.47 (in, 5 H),
7.10-7.18 (in, 2 H), 7.06 (d, Jr.: 9.0 Hz, 2 H),
5.13 (s, 2 H). MS (ES!): calculated for
CI6H14N20, 250.1, found 251.1 [M + H]', 248.8
-
2-(4-Bromopheny1)-1H-itnidazole (91) Yield: 19.5%. 11-1 NMR (300 MHz,
CDC13) 8
12.59 (s, 1 H), 7.87 (d, J= 8.114z, 2 H), 7.64(d,
J-8.1 Hz,! H), 7.27 (s, 1 H), 7.04 (s, 1 H). MS
(ES!) calcd for C91-17BrN2 222.0, found 222.8
+ Hr.
2-(4-(TrifluoromethyDpheny1)-1H-imidazole Yield: 26.2 %; 'H NMR (500 MHz,
CDC13) 3
(91)) 8.03 (d, I = 8.0 Hz, 2 H), 7.66 (d, J= 8.0
Hz, 2
H), '7.25 (s, 2 H). MS (ESL) calcd for C10117F3N2
212.1, found 213.1 [M +H]*.
_
2-(4-nitropheny1)-1H-imidazole (9x) Yield: 53.7 %. 11-1 NMR (500 MHz, DMSO-
d6)
= 8 12.97 (br, 1 H), 8.32 (d, J =9.0 Hz, 2 H), 8.17
(d, J=9.0 Hz., 211), 7.42(s, 1 H), 7.17(s, 111).
MS (ES!): calculated for C9117N302, 189.1,
found 189.9 (M + Hr, 187.8 WI - HT.
161 =
CA 3030689 2019-01-18
2-pheny1-1-(phenylsulfony1)-1H-imidazole Yield: 50.3 %. 1H NMR (500 MHz,
CDC13)
(10a) = 7.64-7.67.(m, 1 H), 7.56 (t, J= 9.0 Hz, 1
H),
7.32-7.48 (m, 9 H), 7.12-7.16 (m, 1 H). MS
(ESL): calculated for C15Hi2N202S, 284.1, found
307.1 [M + Na]4.
2-(4-fluoropheny1)-1-(phenylsulfony1)-1H- Yield: 56.9 %. 'H NMR (500 MHz,
CDC13)
imidazole (10b) 7.66 (d, J= 2.0 Hz, 1 H), 7.58 (1,1 10.0
Hz, 1
H), 7.36-7.42 (m, 6 11), 7.12 (d, J= 2.0 Hz, 1
H), 7.06 (t, I = 10.0 Hz, 2 H). MS (ESO:
calculated for Ci5HliFN202S, 302.1, found
300.8 [M - H].
2-(4-methoxypheny1)-1-(phenylsulfony1)-1H- Yield: 40.9 %. 'H NMR (500 MHz,
CDCI3) 8
imidazole (1(k) 7.62 (d, J= 5.0 Hz, 1 H), 7.56 (n, J-=
15.0 Hz,
5.0 Hz, 1 H), 7.32-7.43 (m, 6 H), 7.10 (d,
5.0 Hz, 1 H), 6.88 (dt,1 .----- 16.0 Hz, 6.0 Hz, 2!-!),
3.87 (s; 3 H). MS (ES!): calculated for
Co61-Ii4N203S, 314.1, found 337.1 [M +Na]4,
312.9 [M -
1-(phenylsulfony1)-2-(p-toly1)-1H-imidazole Yield: 46.6%. 11-I NMR (500
MHz, CDC13) 8
(10d) 7.63 (d, 1= 1.0 Hz, I H), 7.55 (t, J = 8.0
Hz, 1
H), 7.42 (d, I = 8.0 Hz, 2 H), 7.35 (t, J= 7.5 Hz,
2 H), 7.27-7.29 (m, 2 H), 7.16 (d, J= 7.5 Hz, 2
H), 7.10 (s, 1 H), 2.41 (s, 3 H). MS (ES!):
calculated for Ci6H14N202S, 298.1, found 321.1
[M + Nal+.
1-(phenylsulfony1)-2-(3,4,5- Yield: 55.7%. IFI NMR (500 MHz, CDC13)
= tri methoxypheny1)-111-i midazole (10e)
7.68 (d, 1= 1.5 Hz, 1 I-I), 7.55 (t, J= 7.0 Hz, 1
H), 7.42 (d, I = 7.5 Hz, 2 H), 7.35 (1, J= 8.5 Hz,
2 H), 7.11 (d, J= 1.5 Hz, 2 H), 6.60 (s, 1 H),
3.90 (s, 3 H), 3.79 (s, 6 H). MS (ESI): calculated
for Ci811i8N205S, 374.1, found 397.1 [M + Na]4.
2-(4-chloropheny1)-1-(phenylsulfony1)-1H- Yield: 54.9%. 11-1 NMR (500 MHz,
CDC13)
imidazole (101) 7.65 (d, 1= 2.0 Hz, 1 H), 7.58 (t, J= 7.5
Hz, 1
H), 7.43 (d, .1= 8.5 Hz, 2 H), 7.38 (t, J= 8.0 Hz,
2 H), 7.34-7.36 (m, 4 H), 7.12(d, J= 1.5 Hz, 1
H). MS (ES!): calculated for CI511HCIN202S,
318.0, found 341.0 [M + Na].
N,N-dimethy1-4-(1-(phenyl sul fony1)- 1 H- Yield: 48.3%. 111 NMR (300 MHz,
CDC13) 8
imidazol-2-y1) aniline (10g) 7.59 (d, I= 2.0 Hz, 1 11), 7.55 (t, I= 8.0
Hz, 1
H), 7.45 (d, J= 7.5 Hz, 2 H), 7.28-7.38 (m, 4
H), 7.07 (d, J= 2.0 Hz, 1 H), 6.68 (d, J= 8.5
Hz, 2 H), 3.04 (s, 3 II). MS (ESL): calculated for
C17/-117N302S, 327.10, found 350.0 [M + Na]4,
325.9 [M - H].
162
CA 3030689 2019-01-18
4-(1-((4-Methoxyphenyl)sulfony1)-1H- Yield: 61.5%. 'H NMR (500 MHz, CDC13)
irnidazol-2-y1)-N,N-dimethylaniline (10gb) 7.58 (d, J = 1.5 Hz, 1 H), 7.36
(t, J = 8.43 Hz,
4 H), 7.03 -7.09 (m, 1 H), 6.80 (d, J = 9.0 Hz,
2 H), 6.69 (d, J = 8.8 Hz, 2 H), 3.84 (s, 3 H),
3.05 (s, 6 H). MS (ESI): calculated for
Ci7Hi7N302S, 327.1, found 358.2 [M + Nar.
2-(3,4-dimethoxypheny1)-1-(phenylsulfony1)- Yield: 60.3%. 11-1 NMR (500
MHz, CDC13)
(HA midazole (10h) 7.64 (d, J -= 7.0 Hz, 1 H), 7.55 (t, I =
7.5 Hz, 1
H), 7.40 (dd, J = 8.5 Hz, 1.5 Hz, 2 H), 7.35 (t, J
= 8.0 Hz, 211), 7.09 (d, J= 2.0 Hz, I H), 7.02
(dd, J = 8.0 Hz, 2.0 Hz, I H), 6.89 (d, J = 1.5
Hz, I H), 6.86 (d, J = 8.0 Hz, 1 H), 3.95 (s, 3
HI 3.81 (s, 3 H). MS (ESI): calculated for
CI7H16N204S, 344.10, found 367.0 [M + Na].
1-(phenylsulfony1)-2-(2- Yield: 58.6%. 'H NMR (500 MHz, CDC13) 8
(trifluoromethyl)pheny1)-1H-imidazole (1W) 7.64-7.67 (m, 2 H), 7.61-7.63
(m, 3 H), 7.40-
7.46 (m, 5 H), 7.16 (d, J = 1.5 Hz, 1 H). MS .
(ESI): calculated for C161-111F3N202S, 352.10,
found 353.1 [M + Hr.
2-(4-(benzyloxy)pheny1)-1-(phenylsulfony1)- Yield: 62.0%; mp 102 - 104 C.
114 NMR (500
1H-imidazole (10j) MHz, CDC13) 67.56 (d, J= 1.0 Hz, 1 H),
7.46
(t, J = 8.0 Hz, 1 H), 7.20-7.40 (m, 11 H), 7_03
(d, J = 1.0 Hz, 1H), 6.89 (t, J = 8.0 Hz, 2 H),
5.08 (s, 2 H). MS (ESI): calculated for
C22H18N203S, 390.10, found 413.1 [M + Na]4.
HPLC2: tit 18.22 min, purity 95.9%.
2-(4-Bromopheny1)-1-(phenylsul fony1)-1 PI- Yield: 61.2%. 1H NMR (500 MHz,
CDC13) 8
imidazole (101a) 7.71 (d, J = 2.0 Hz, 1 H), 7.64 (t, J =
7.0 Hz, I
H), 7.57 (d, I = 9.0 Hz, 2 H), 7.49 (d, J = 7.0
Hz, 2 H), 7.45 (t, J = 9.0 Hz, 2 H), 7.34 (d, J =
8.5 Hz, 2 H), 7.18 (d, J = 1.5 Hz, 1 H). MS
(ESI) calcd for C151-111BiN202S 362.0, found
363.0 [M + Hr.
-(Phenylsulfony1)-2-(4- Yield: 36.7 %; 114 NMR (500 MHz, CDC13)
(VW uoromethyl)pheny1)-1H-imidazole (10p) 7.75 (d, J = 2.0 Hz, 1 H), 7.69
(d, J = 8.0 Hz, 2
H), 7.65 (t, J = 8.0 Hz, 1 H), 7.60 (d, J= 8.0 Hz,
2 H), 7.48 (d, I = 7.5 Hz, 2 11), 7.43 (t, J = 8.0
Hz, 2 H), 7.22 (d, J = 2.0 Hz, 1 H). MS (ESI)
calcd for Ci6H1IFIN202S 352.1, found 553.1 f111
+
2-(4-nitropheny1)-1-(phenylsulfony1)-1H- Yield: 50%; mp 145 - 147 C. 'H
NMR (500-
irnidazole (10x) MHz, DMSO-d6) 6 8.28 (d, J =8.5 Hz, 2 H),
8.03 (d, J = 1.5 Hz, 1 H), 7.78 (t, J = 7.5 Hz, 1
H), 7.64-7.68 (m, 4H), 7.60 (t, J = 8.0 Hz, 2 H),
7.30 (d, I = 1.5 Hz, 1 H). MS (ESI): calculated
for C151-111N304S, 329.10, found 352.0 [M +
Nar, 327.9 [M - FIT. HPLC2: ri/ 14.87 min,
_purity 98,8%.
163
CA 3030689 2019-01-18
(4-methoxyphenyl)(2-phenyl-1- Yield: 26.3%; mp 118 - 120 C. NMR (500
(phenylsulfony1)-1H-imidazol-4-yl)methanone MHz, DMSO-d6) 8 8.37 (d, J =
1.0 Hz, 1 HI
(11ab) 8.15-8.18 (m, 2 H), 8.12 (d, J = .9.0 Hz,
2 H),
7.56-7.64 (m, 5 H), 7.46-7.50 (m, 3 H), 7.16 (d,
J = 8.0 Hz, 2 H), 3.90 (s, 3 H). MS (ES1):
calculated for C23111BN204S, 418.10, found
419.1 [M + H]t. HPLC2: tR 17.72 min, purity
95.7%.
(3- methoxyphenyl)(2-phenyl- 1- Yield: 31.2%; mp 136- 138 C. '1-1 NMR
(500
(phenyIsulfony1)-1/1-imidazol-4-yl)methanone MHz, CDC13) 8 8.35 (s, 1 H), 7.86
(d, J = 8.0
(1 lac) Hz, 1 H),7.72 (s, 1 H), 7.60(t, J=7.5 Hz,
1 H),
7.51 (t, J = 7.5 Hz, 1 H), 7.35-7.42 (m, 911),
7.14 (dd, J = 8.0 Hz, 2.0 Hz, 1 H), 3.88 (s, 3 H).
MS (ES1): calculated for C231118N204S, 418.10,
found 419.1 [M + HPLC2: tR
.17.72 min,
_purity 95.7%.
(2-pheny1-1-(phenylsulfony1)-1H-imidazol4- Yield: 28.9%; mp 108 - 110 C. H
NMR (500
yl)(p-tolyl)methanone (11ah) MHz, CDC13) S 8.00 (d, J = 7.5 Hz, 2 H),
7.98
(q, J = 8.0 Hz, 1.5 Hz, 2 H), 7.91 (d, J = 8.0 Hz,
1 H), 7.81 (s, 1 H), 7.44-7.48 (m, 3 H), 7.35-
7.40 (m, 2 H), 7.30 (t,J = 8.0 Hz, 2 H), 7.20 (s,
2 H), 2.42 (s, 3 H). MS (ESI): calculated for
C23HiaN203S, 402.10, found 403.1 [M + Hr.
HPLC2: tR 16,06 min, purity 96.2%.
(4-fluorophenyl)(2-phenyl-1-(phenylsulfony1)- Yield: 25.4%; mp 114- 116 C.
IF1 NMR (500
1H-imidazol-4-yl)methanone(llaf) MHz, CDC13) 8 8.10 (q, J = 3.5 Hz, 5.5 Hz,
2
H), 7.88 (d, J = 7.5 Hz, 2 11), 7.67 (t,J= 7.5 Hz,
1 H), 7.48 -7.54 (m, 3 H), 7.38- 7.41 (m, 5 H),
7.24 (t, J = 8.5 Hz, 2 H). MS (ESI): calculated
for C221115FN203S, 406.10, found 429.1 [M +
Na]t. HPLC2: tR 15.43 mm, punt 96.1%.
(3-fluorophenyl)(2-phenyl-1-(phenylsulfony1)- Yield: 18.3%; mp 102 - 104 T. H
NMR (500
'HA midazol-4-yl)methanone(llag) MHz, CDC13) 8 8.14 (d,J = 7.5 Hz, 1 H),
7.76
7.87 (m, 3 H), 7.74 (d, J = 9.0 Hz, 1 H), 7.37 -
7.57 (m, 10 H), 7.38 - 7.41 (m, 5 H), 7.24 (t,.) =
8.5 Hz, 2 H). MS (PSI): calculated for
C22H,3FN,03S, 406.10, found 429.1 [ivl + Na]t,
HPLC2: tR 15.75 min, purity 96.5%.
= (4-fluorophenyl)(2-(4-metimxyphen y1)-1- Yield: 23.5%; mp 135 -
137 C. 11-1 N1V1R (500
(phenylsulfony1)-111-imidazol-4-ypmethanone MHz, CDC13) 8 8.00 (d,J= 5.5 Hz, 2
H), 7,74 -
(110) 7.76 (m, 2 H), 7.54-7.58 (m, 1 H), 7.40
(d, J =
7.0 Hz, 2 H), 7.28-7.30 (m, 3 H), 7.14- 7.16(m,
2 H), 6.80-6.82 (m, 2 H), 3.80 (s, 3 H). MS
(ESD: calculated for C231-117FN204S, 436.10,
found 459.0 TM + Nar, 434.9 [M - Hr.
HPLC2: tR 16.53 min, Purity 96.1%.
164
CA 3 0 3 0 6 8 9 2 0 1 9-0 1-1 8
(1-(phenylsulfony1)-2-(p-to1y1)-1H-imida7o1- Yield: 33.8%; 111 NMR (500
MHz, CDCI3)
4-y1)(3,4.5 -tri methoxyphenyHmethanone 8.00 (d, J= 8.0 Hz, 2 H), 7.70 (t,
J= 7.5 Hz, 1
(11da) H), 7.55 (t, )= 8.0 Hz, 2 H), 7.44 (s, 2
H), 7.34
(s, 2H), 7.31 (d, J= 8.0 Hz, 2 H), 7.21 (d, J=
8.0 Hz, 2 H), 4.00 (s, 3 H), 3.98 (s, 6 H). MS
(ES!): calculated for C261-124N206S, 492.14, '
found 515.2 [M + Na].
(4-fluorophenyl)(1-(phenylsulfony1)-2-(p- - Yield:
18.6%; mp 142 - 144 C. 'El NMR (500
toly1)-1H-imidazol-4-yl)methanone (11db) MHz, CDCI3) 8 8.07 (q, J= 8.5 Hz,
5.5 Hz, 2
H), 7.88 (d, J =7.5 Hz, 2 1-0, 7.64 (t, J = 8.0 Hz,
1 H), 7.49 (d, J = 8.0 Hz, 2 H), 7.38 (s, 1H),
7.30 (d, I= 8.0 Hz, 2 H), 7.18 - 7.24 (m, 4 H),
2.43 (s, 3 H). MS (ES!): calculated for
Ci3Hi7EN203S, 420.10, found 443.0 [M + Nar,
418.9 [M - HPLC2: 111
17.28 min, purity
97.3%.
(1-(phenylsulfony1)-2-(3,4,5- Yield: 21.1%; mp 135- 137 C. '11 NMR (500
trimethoxypheny1)-1H-imidazol-4-y1)(3,4,5- MHz, CDC13) 37.91 (d, J=8.0 Hz,
2 H), 7.65 (t,
trimethoxyphenypmethanone (ilea) J= 7.5 Hz, 1 H), 7.51 (t, J= 8.0 Hz, 2 H),
7.44
(s, 1 H), 7.34 (s, 2 H), 6.60 (s, 2 H), 3.98 (s, 3
H), 3.96 (s, 5 H), 3.91 (s, 3 H), 3.73 (s, 6 H).
MS (ES!): calculated for C2s1-12,3N209S, 568.2,
found 569.2 [M + H]. HPLC1: IR 17.86 min,
purity 98.9%.
(4-fluorophenyl)(1-(phenylsulfony1)-2-(3,4,5- Yield: 18.8%; mp 135 - 137
C. 'H NMR (500
trimethoxypheny1)-1H-i midazol-4- MHz, CDC13) 5 8.11 (q. J =5.5 Hz, 3.0 Hz,
1
yl)methanone (1 leb) H), 8.00 -8.03 (m, 1 H), 7.82 (d, 1= 7.5
Hz, 1
H), 7.78 (s, 1 H), 7.64 (t, J =7.0 Hz, 1 H), 7.48
(t, J =8.0 Hz, 1 1-1), 7.42 (s, 1 H), 7.21 -7.26 (m,
4 HI 6.62 (s. 1 H), 3.98 (s, 3 H), 3.96 (s. 6 H),
3.93 (s, 3 H). MS (ES!): calculated for
C251-121FN206S, 496.10, found 497.1 [M + Hr.
HPLC2: rit 15.26 min, purity 98%.
(2-(4-chloropheny1)-1-(phenylsulfony1)-1H- Yield: 36.8%; mp 153 - 155 C.
'H NMR (500
imidazol-4-y1)(4-fluorophenyl)methanone MHz, CDCI3) 5 8.0&(q, J =5.5 Hz,
3.0Hz, 2 H),
(11f13) 7.89 (d, J =7.5 Hz, 2 H), 7.68 (t, J =8.0
Hz, 1
H) , 7.52 (t, J = 8.0 Hz, 2 H), 7.34-7.38 (m, 5H),
7.23 (t, J =8.5 Hz, 2 H). MS (EST): calculated
for C221114CIFN203S, 440.0, found 463.0 [M +
Na]. HPLC2: rit 17.72 min, purity 97.38%.
(2-(4-(di me th yl ami n o)ph e ny1)-1- Yield: 32.2%; mp 157 - 159 C. 11-1
NMR (500
(phenylsulfony1)-1H-i midazol -4-y1)(3,4,5- MHz, CDC13) 67.89 (d, J=8.0 Hz,
2 H), 7.62 (t,
trimethox yphenyemethanone (11ga) J=7.5 Ilz, 1 1-1), 7.48 (t, J =8.0 Hz. 2
H), 7.43
(s, 1 H), 7.32 (d, J=8.5 Hz, 2 H), 7.30 (s, 2H),
6.62 (d, J=9.0 Hz, 2 H), 3.97 (s, 3 H), 3.95 (s, 6
H), 3.05 (s, 6 H). MS (ESI): calculated for
C27I127N306S, 521.2, found 544.1 [M + Na],
519.8 [M - H]. HPLC2: IR 16.00 min, purity
97.9%.
165
CA 3030689 2019-01-18
(2-(4-(dimethylamino)phenyI)-1- Yield:
38.5%; mp 125 - 127 C. 'H NMR (500
(phenylsulfony1)-1H-imidazol-4-y1)(4- . MHz,
CDC13) 6 8.04 (q, =5.5 Hz, 3.5Hz, 2 H),
fluorophenypmethanone (11gb) 7.80 (d, J
=7.5 Hz, 2 H), 7.61 (t, J =8.0 Hz, 1
H), 7.45 (t, 1=8.0 Hz, 2 H), 7.39 (s, 1 H), 7.35
(d,19.0 Hz, 211), 7.21 (t, =8.5 Hz, 2 H), =
6.62 (d, J =9.0 Hz, 2 H), 3.05 (s, 6 H). MS
(ESI): calculated for C241-120FN303S, 449.10,
found 472.1 [N4 + Na], 447.9 [M - 111".
HPLC2: tR 16.85 min, purity 96.5%.
(2-(3,4-dimethoxypheny1)-1-(phenylsulfony1)- Yield: 28.6%; mp 136- 138 C. 'H
NMR (300
1H-imidazol-4-y1)(3,4,5- MHz, CDC13)
6 7.92 (dd, J =8.5 Hz, 1.5 Hz, 2
trimethoxyphenyl)methanone (11ha) H), 7.66 (t,
J=7.5 Hz, 2 H), 7.51 (t, J=7.5 Hz, 2
H), 7.43 (s, 1 H), 7.33 (s, 2 H), 7.02 (dd, J =8.0
Hz, 2.0 Hz, 1 H), 6.91 (d, J =2.0 Hz, 1 H),6.86
(d, J =8.5 Hz, 1 H), 3.98 (s, 3 H), 3.96 (s, 9 H),
3.77 (s, 3 H). MS (ES!): calculated for
C271126N2O8S, 538.10, found 561.1 [M + Nar,
536.8 (M - Hr. HPLC2: tR 14.67 min, purity
98.2%.
(2-(3,4-dimethoxypheny1)-1-(phenylsulfonyl)- Yield: 31.9%; mp 144- 145 C. 11-
1 NMR (300
1H-imidazol-4-y1)(4-fluorophenyl)methanone MHz, CDC13) 3 8.09 (q, I =5.5 Hz,
3.5 Hz, 2
(11hb) H), 7.81 (d, J =8.0 Hz, 2 H), 7.62 (t, J =7.5
Hz, 2 H), 7.48 (t, J =7.5 Hz, 2 H), 7.40 (s, 1
H), 7.21-7.25 (m, 2 H), 7.04 (dd, J =8.0 Hz,
2.0 Hz, 1 H), 6.92 (d, J =2.0 Hz, 1 H), 6.86 (d,
J =8.5 Hz, 1 H),3.96 (s, 3 H), 3.79 (s, 6 H).
= MS (ES!): calculated for C241119FN205S,
466.10, found 489.1 [M + Na]', 464.8 [M -
H]. HPLC2: tR 15.52 mm, purity 97.4%.
(1-(phenylsulfony1)-2-(2- Yield:
25.0%; mp 155 - 157 C. 111 NMR (500
(trifluoromethyl)pheny1)-1H-i midazol-4- MHz, DMSO-
d6) 8 7.91 (d, J =8.0 Hz, 1 H),
yl)(3,4,5-trinnethoxyphenyl)methanone (Ilia) 7.84 (q,
=7.5 Hz, 5.0 Hz, 211), 7.77-7.80(m, 2
H), 7.75 (s, 2 H), 7.66 (t, J =8.0 Hz, 2 H), 7.56
(d, 1 =7 .5 Hz, 1 H), 7.18 (s, 2 H), 3.87 (s, 6 H),
3.81 (s, 3 H). MS (ES!): calculated for
C26H21F3N206S, 546.10, found 569.0 [M + Nal+.
HPLC2: rit 16.16 min, purity 98.9%.
(1-(phenylsulfony1)-2-(2- Yield:
25.0%; mp 151 - 153 C. 11-1 NMR (500
(trifluoromethyl)pheny1)-1H-itnidazol-4-y1)(4- MHz, CDC13) 8 8.03 (q, J =5.5
Hz, 3.0 Hz, 2
fluorophenypmethanone (llib) H), 7.90 (d, J =8.0 Hz, 2 H), 7.80 (d, J
=8.0 Hz,
1 H), 7.69 (q, J =7.0 Hz, 6.5 Hz, 2 11), 7.61 (t, J
=8.0 Hz, 1 H), 7.52 (t, J =8.0 Hz, 2 H), 7.34 -
7.36 (in, 2 H), 7.23 (t, J =8.5 Hz, 2 H). MS
(ES!): calculated for C231114F4N203S, 474.10,
found 497.0 [N4 + Na]'. HPLC2: tR 16.80 min,
purity 98.2%.
166
CA 3030689 2019-01-18
(2-(4-(benzyloxy)pheny1)-1-(phenylsulfony1)- Yield: 22.3.0%; mp 149 - 151 C.
'H NMR
1H-imidazol-4-y1)(4-fluorophenyflmethanone (500 MHz, CDC13) 8 8.09 (q, J =5.5
Hz, 3.5 Hz,
(11jb) 2 11), 7.82 (d, J =7.5 Hz, 2 H), 7.63 (t,
7.5 Hz, 1
H), 7.36-7.50(m, 10 H), 7.25 (t, J =8.5 Hz, 2 H),
6.98 (d, J =8.0 Hz, 2 H), 5.17 (s, 2 H). MS
(ESI): calculated for C291-121EN204S, 512.10,
found 535.0 [M + Nat. HPLC2: tit 18.35 min,
purity 95.1%.
(2-(4-bromopheny1)-1-(phenylsulfony1)-111- Yield: 32.6% 1H NMR (500 MHz,
CDC13)
inidazol-4-y1)(3,4,5- 8.06 (d, J = 8.0 Hz, 2 H), 7.88 (d, J =
8.5 Hz, 1
trimethoxyphenyl)methanone (111a) H), 7.77 (t, J = 7.0 Hz, 1 1-1), 7.54-
7.63 (m, 4
H), 7.31-7.36 (m, 4 H), 4.04 (s, 3 H), 4.01 (s, 6
H). MS (ESI) calcd for C25H2113rN206S 556.0,
found 557.0 [M +
(1-(phenylsulfony1)-2-(4- Yield: 36.7 %; IF1 NMR (500 MHz, CDC13) 8
(trifluoromethyl)pheny1)-1H-i midazol-4- 8.06 (d, I = 7.5 Hz, 2 H), 7.78
(t, J = 8.0 Hz, 1
= yl)(3,4,5-trimethoxyphenyl)methanone (11pa) H), 7.72 (d, J = 8.0 Hz, 2
H), 7.62 (d, J = 8.0
Hz, 2 H), 7.59 (d, J = 8.0 Hz, 2 H), 7.50 (s, 1
H), 7.37 (s, 2 1-1), 4.04 (s, 3 H), 4.02(s, 6 H).
MS (ESI) calcd for C26H2IP3N206S 546.1,
found 547.11M + Hr.
(2-(4-(dimethylamino)phenyl)-1-04- Yield: 34.1%; mp 147-149 C. 11-1 NMR
(500
methoxyphenypsulfony1)-1H-imidazol-4- MHz, CDC13) 8 8.07 (q, J = 8.5 Hz,
5.5 Hz, 2
yl)(3,4,5-trimethoxyphenyflmethanone FI), 7.78 (d, J = 9.0 Hz, 2 H), 7.41
(d, J = 8.5
(11gaa) Hz, 2 H), 7.39 (s, 1 H), 7.23 (t, J = 8.5
Hz, 2
H), 6.91 (d, J = 9.0 Hz, 2 H), 6.68 (d, J = 9.0
Hz, 2 H), 3.89 (s, 3 H), 3.08 (s, 3 H). MS
(ESI) calcd for C281-12gN301S 551.2, found
573.1 [M + Nar. HPLC2: tR 18.6 min, purity
96.9%.
(2-phenyl- I H-i midazol -4-y1)(3,4,5- Yield: 10.1 %; mp 227-229 C. 'H NMR
(500
trimethoxyphenyflmethanone (12aa) MHz, CDC13) 5 8.0-8.03 (m, 2 H), 7.83 (s,
1 H),
7.34-7.38 (m, 3 H), 7.21 (s, 2 H), 3.90 (s, 3 H),
3.84 (s, 6 H). MS (ESI): calculated for
Colli8N20, 338.1, found 337.1 [M - H]
HPLC2: tg14.19 min, purity 96.3%.
(4-methoxyphenyl)(2-phenyl-1H-imidazol-4- Yield: 16.6%; mp 179 - 181 C. 1H
NMR (500 =
ypmethanone (12ab) MHz, CDC13) 8 11.1 (br, 1 H), 8.07-8.10
(m, 2
H), 8.04 (d, J = 8.5 Hz, 2 11), 7.84 (d, J = 1.0
Hz, I H), 7.49-7.51 Cm, 3 H), 7.07 (d, J = 9.0
Hz, 2 H), 3.95 (s, 3 H). MS (ESL): calculated for
CI7H14N202, 278.10, found 279.0 [IV + Hr.
HPLC I : tR 15.14 min, purity > 99%.
167
CA 3030689 2019-01-18
(3-methoxyphenyl)(2-phenyl-1H-imidazol-4- Yield: 22.5%; mp 160 - 162 C. 'H
NMR (500
yl)methanone (12ac) MHz, CDCI3) 511.2 (br, 1 H), 8.10-8.12
(in, 2
H), 7.87 (d, J = 1.0 Hz, 1 H), 7.61 (d, I = 7.5
Hz, 1 H), 7.48- 7.52 (m, 5 H), 7.21 (dd, J = 2.5
Hz, 8.5Hz, 1 H), 3.91 (s, 3 H). MS (ES!):
calculated for Co1114N202, 278.10, found 279.0
[M + Hr. HPLC2: tR 15.07 min, Tufty >99%.
(3,5-dimethoxyphenyl)(2-phenyl-1H- Yield: 26.2%; mp 168- 170 C. H NMR (500
imidazol-4-yl)methanone (12ad) MHz, CDCI3) 8 8.04-8.06 (in, 2 H), 7.88
(s, 1
H), 7.50-7.52 (m, 3 H), 7.15 (d, J = 2.0 Hz, 2
H), 6.75 (t, J = 1.0 Hz, 1 H), 3.89 (s, 6 H). MS
(ES!): calculated for C1s1116N203, 308.10, found
331.1 [M + Na}, 306.9 [M - H]. HPLC2: tit
15.59 min, purity > 99%.
(3.4-dimethoxyphenyl)(2-phenyl-1H- Yield: 18.6%; mp 162 - 164 C. 111 NMR
(500
iinidazol-4-Amethanone (12ae) MHz, CDCI3) 8 10.9 (br, 1 H), 8.05 (dd, J
= 1.5
Hz, 8.0 Hz, 2 H), 7.86 (d, J= 1.5 Hz, 1 11), 7.74
(dd, J = 2.0 Hz, 8.5 Hz, 1 H), 7.56 (d, J = 2.0
Hz, 1 H), 7.50-7.52 (m, 3 H), 7.04 (d, J = 8.5
Hz, I H), 4.03 (s, 3 H), 3.99 (s, 3 H). MS (ES!):
calculated for C181-116N203, 308.10, found 331.1
[Is4 + Na], 306.9 - HPLC2: tit
13.54
min, purity > 99%.
(4-fluorophenyl)(2-phenyl-1H-imidazol-4- Yield: 30.2%; mp 231 -233 C. 'H
NMR (500
yOmethanone (12af) MHz, CDC13) 5 10.6 (br, 1 H), 8.02-8.05
(m, 4
H), 7.81 (d, J = 1.0 Hz, 1 H), 7.51-7.54 (m, 3
H), 7.27 (t, J = 8.5 Hz, 2 11). MS (ESI):
calculated for Ci6111,FN20, 266.10, found 267.0
= [M +H].,
264.8 [M HPLC I: tR 15.37 mm,
purity 98.9%.
(3-fluorophenyl)(2-phenyl-IH-irnidazol-4- Yield: 23.4%; mp 212- 214 C. 111
NMR (500
yl)methanone (12ag) MHz, CDCI3) 8 8.05 (dd, J = 1.5 1-1z, 7.5
Hz, 2
H), 7.86 (s, 1 H), 7.84 (d, J = 7.0 Hz, 1 H), 7.74
(d, J = 8.5 Hz, 1 H), 7.52-7.58 (m, 4 H), 7.37
(dt, J =2.0 Hz, 6.0 Hz, 1 H). MS (EST).
calculated for CI6HilFN20, 266.10, found 267.0
[M +H], 264.8 [M - Hr. HPLC 1: tit 15.29 min,
purity > 99%.
(2-phenyl-1H-imidazol-4-y1)(p- Yield: 15.6%; nip 225 - 227 C. NMR
(500-
tolyl)methanone (12ab) MHz, CDCI3) 5 11.1 (br, 1 H), 8.08 (d, J =
7.5
Hz, 2 II), 7.93 (d, J =9.0 Hz, 2 H), 7.84 (s, 1 H),
7.48-7.52 (m, 3 H), 7.38 (d, J = 10.0 Hz, 2 H),
2.50 (s, 3 H). MS (ESL): calculated for
' CI7H141420, 262.10, found 263.0 [M
260.8 [M - H]. HPLC2: IR 15.86 min, purity
98.7%.
=
168
CA 3030689 2019-01-18
(2-phenyl-1H-imidazol-4-y1)(m- Yield: 20.5%;
mp 168 - 169 C. '11 N1V1R (500
tolypmethanone (12ai) MHz, CDC13) 5
11.0 (br, 1 H), 8.09-8.11 (m, 2
El), 7.84 (d, J = 1.5 Hz, 1 H), 7.81-7.82 (m, 2
H), 7.47-7.52 (m, 5 H), 2.50 (s, 3 H). MS (ES!):
calculated for C171114N20, 262.10, found 285.0
[M +Nar, 260.8 [M - H]. HPLC2: tR 15.89
, min, purity >99%.
(2-(4-fluoropheny1)-1H-imidazol-4-y1)(3,4,5- Yield: 12.2%.
mp 176- 178 C. NMR (500
trimethoxyphenyflmethanone (12ba) MHz, CDC13) 5 10.72 (br, 1 H), 8.02
(q, J = 5.0
Hz, 2 H), 7.84 (s, 1 H), 7.19 (t, J = 10.0 Hz, 2
H), 4.00 (s, 6 H), 3.97 (s, 3 I-1). MS (F51):
calculated for Ci9Hi7FN204, 356.10, found
379.1 [M + Na], 354.9 [M - HI. HPLC1: tR
17.23 min, purity > 99%
(2-(4-methoxypheny1)-1H-i midazol-4- Yield: 10.2%;
nip 220 - 222 C. Ili NMR (300
yl)(3,4,5-trimethoxyphenypmethanonc (12ca) MHz, CDCI3) 5 10.24 (br, 1 H), 7.93
(d, J =
14.5 Hz, 2 H), 7.81 (s, 1 H), 7.24 (s, 2 H), 7.03
(d, J = 14.5 Hz, 2 H), 3.97 (s, 3 H), 3.95 (s, 6
H), 3.90 (s, 3 H). MS (ES!): calculated for
C2,3E120N205, 368.10, found 391.0 [M + Na]4,
367.0 [M - HPLC2: IR
14.46 min, purity
98.4%.
(4-fluorophenyl)(2-(4-methoxypheny1)-1H- Yield: 15.2%;
mp 245 - 247 C. NMR (500
iruidazol-4-ypmethanone (12cb) MHz, CDC13) 8 10.20 (br, 1 H), 7.93-7.96
(m, 2
H), 7.85 (d, 3 5.0 Hz, 2 H), 7.68 (s, I H), 7.15-
7.17 (m, 2 H), 6.95 (d, J= 6.0 Hz, 21-!), 3.82(s,
3 H). MS (E51): calculated for Ci7F113FN202,
296.10, found 319.1 [M + Na]4, 294.9 [M - Hr.
HPLC2: tR 15.40 min, purity 98.8%.
(2-(p-toly1)-1H-imidazol-4-y1)(3,4,5- Yield: 48.5%;
mp 201 -203 C. NMR (500
trimethoxyphenypmethanone (12da) MHz, CDC13) 8 10.40 (br, 1 H), 7.88
(d, J = 8.0
Hz, 2 H), 7.82 (s, 1 H), 7.31 (d, J = 8.0 Hz, 2
H), 7.24 (s, 2 H), 3.96 (s, 3 H), 3.94 (s, 6 H),
2.43 (s, 3 H). MS (EST): calculated for
C201120N204, 352.10, found 375.2 [M + Na]4.
HPLC2: tR 15.45 min, purity 97.4%.
(4-fluorophenyl)(2-(p-toly1)-1H-imidazol-4- Yield: 56.3%;
up 229 - 231 C. NMR (500
yl)methanone (12db) MHz, CDC13) 8
10.50 (br, 1 H), 7.99-8.02 (m, 2
H), 7.88 (d, J 8.0 Hz, 2 H), 7.60 (d, J = 1.0
= Hz, I H), 7.30 (d, J 8.0 Hz, 2 11), 7.23 (t,
9.0 Hz, 2 H), 2.43 (s, 3 H). MS (ES!): calculated
for CHH13FN20, 280.10, found 281.0 [M +
278.9 1M - Hr. HPLC2: tR 16.31 mm, purity >
99%.
(4-hydroxy-3,5-dimethoxyphertyl)(2-(p-toly1)- Yield: 56.8%; mp 220-222 C. 'H
NMR (500
1H-imidazol-4-yl)methanone (12de) MHz, CDC13) 8 8.02 (d, J = 8.0 Hz,
211),
7.91(s, 111), 7.39 (s, 2H), 7.28 (d, J = 7.5 Hz,
2H), 4.00 (s, 6H), 2.44 (s, 3H). MS (PSI) calcd
for C19Hi8 PN204 338.1, found 339.1 [M +
H]4. ELPLCI: tR 3.91 min, purity > 99%.
169
CA 3030689 2019-01-18
(3,4,5-trimethox yphenyl)(2-(3,4,5- Yield: 86.8%; mp 196 - 198 C. 'H NMR
(500
trimethoxypheny1)-1H-i midazol-4- MHz, DMSO-d6) 5 13.3 (br, 0.47 H), 13.50
(br,
yl)methanone (12ea) 0.52 H), 8.19 (s, 0.49 H), 7.90 (s, 1 H),
7.83 (s,
0.5 H), 7.59 (s, 1 H), 7.40 (s, 1 HI 7.18 (s, 1 H), =
3.89 (s, 6 H), 3.86 (s, 6 H), 3.77 (s, 3 H), 3.72 (s,
3 H). MS (ES!): calculated for C22H24N207,
428.2, found 451.1[M + Nal+, 426.9 [M - Hr.
HPLC2: tR 14.49 min, purity >99%.
(4-fluorophenyl)(2-(3,4,5-trimethoxypheny1)- Yield: 90.2%; mp 153 - 155 C. 'H
NMR (500-
1H-i midazol-4-y1)methanone (12eb) MHz, CDC13) 5 10.42 (br, 1 H), 8.00 (q,
J = 5.5
Hz, 3.0Hz, 2 H), 7.76 (s, 1 H), 7.23 (t, J = 8.5
= Hz, 2 1-0, 7.19 (s, 2 H), 3.94 (s, 3 H), 3.92 (s, 3
H). MS (ES!): calculated for C191-117EN204,
356.1, found 379.0 [M + Na], 354.9 [M - Hr.
HPLC2: tR 15.31 min, purity >99%.
(2-(4-chloropheny1)-1H-imidazol-4-y1)(3,4,5- Yield: 36.9%; mp 193 - 195 "C.
11-1 NrvIR (500
trimethoxyphenyl)methanone (12fa) MHz, CDC13) 5 10.75 (br, 1 H), 7.96 (d, J
= 8.5
Hz, 2 H), 7.83 (s, I H), 7.47 (d, J = 9.0 Hz, 2
H), 7.23 (s, 2 H), 3.97 (s, 3 H), 3.94 (s, 6 H),
2.43 (s, 3 H). MS (ES!): calculated for
C191-117C1N204, 372.1, found 395.1 [M + Na},
370.9 [M -Hr. HPLC2: tR 16.36 min, purity >
99%.
(2-(4-chloropheny1)-1H-imidazol-4-y1)(4- Yield: 83.7%; mp 232 - 234 C. 'H
NMR (500
fluorophenyl)methanone (12Th) MHz, CDC13) 5 10.78 (br, 1 H), 8.00 (q,
1=5.5
Hz, 3.0Hz, 2 H), 7_96 (d, J = 9.0 Hz, 2 H), 7.78
(s, 1 H), 7.47 (d, J = 8.0 Hz, 2 H), 7.24 (t, J =
8.5 Hz, 2 H). MS (ES!): calculated for
C161110C1FN20, 300.1, found 323.0 [M + Nar,
298.8 [M HT. HPLC2: tR 17.08 min, purity >
99%.
(2-(4-chloropheny1)-1H-i midazol -4 -y1)(4- Yield: 80.2%; mp
216-218 'C. NMR (500
= hydroxy-3,5-dimethox ypheny Dmethanone
MHz, CD30D) 5 8.06 (d, J = 8.5 Hz, 2 H), 7.99
(12fc) (s, 1 H), 7.61 (d, J = 8.0 Hz, 2 H), 7.52
(s, 2 H),
4.01 (s, 6 H). MS (ESI) calcd for CI8HI5C1N204
358.1, found 359.1 [M + Hr. HPLC2: tR 4.12
min, purity > 99%.
(2-(4-(dimethylamino)pheny1)-1H-imidazol-4- Yield: 91.2%; mp 195 - 197 C. 11-
1 NMR (500
yl)(3,4,5-trimethoxyphenyl)methanone (12ga) MHz, CDC13) 8 10.39 (br, 1 H),
7.87 (d, I = 8.5
Hz, 2 H), 7.80 (s, 1 H), 7.23 (s, 2 H), 6.75(d, 1=
9.0 Hz, 2 H), 3.95 (s, 3 FO, 3.94 (s, 6 /I), 3.05 (s,
6 H). MS (ES!): calculated for C211-123N304,
381.2, found 404.2 [M + Nar, 380.0 [M - H].
HPLC2: tR 15.20 min, purity 95.8%.
170
CA 3030689 2019-01-18
(2-(4-(dimethylamino)pheny1)-1H-imidazol-4- Yield: 86.7%; mp 278 - 280 C. 11-
1 NMR (500
yl)(4-fluorophenyl)methanone (120) MHz, CDC13-) 5 10.21 (br, 1 H), 7.98 (q,
J = 5.0
Hz, 3.5Hz, 2 HI 7.84 (d, J 8.5 Hz, 2 H), 7.72
(s, 1 H), 7.20 (t, J = 8.5 Hz, 2 H), 6.76 (t, J =9.0
= Hz, 2 H), 3.06 (s, 6 H). MS (ESI): calculated for -
C181116FN30, 309.1, found 332.1 [M +
307.9 (M - H. HPLC2: tR 16.06 min, purity
95.6%.
(2-(3,4-di methox ypheny1)- midazol-4- Yield: 85.0%; mp 100- 102
C. 'H NMR (500
yl)(3,4,5-trimethoxyphenyl)methanone (12ha) MHz, CDC13) 5 10.19 (br, 1 H),
7.81 (s, I fl),
7.58 (d, Jr 1.5 Hz, 1 H), 7.48 (d, J = 8.0 Hz, 1
H), 7.25 (s, 2 H), 6.97 (d, J = 8.5 Hz, 1 H), 4.00
(s, 3 H), 3.96 (s, 6 H), 3.95 (s, 6 H). MS (ESI):
calculated for C211422N206, 398.2, found 399.1
[M + Hr, 397.0 [M - HT. HPLC2: k 13.73
mm, purity > 99%.
(2-(3,4-dimethoxypheny1)-1H-imidazol-4- Yield: 78.3%; mp 174- 176 C. 11-1
NMR (500
yl)(4-fluorophenyl)methanone (12hb) MHz, CDC13) 5 8.02 (t, J = 9.0 Hz, 2 HI
7.75
(s, 1 H), 7.57 (s, 1 H), 7.48 (d, J 8.5 Hz, 1 H),
7.23 (t, J = 8.5 Hz, 2 H), 6.95 (d, J = 8.5 Hz, 1
H), 3.99 (s, 3 H), 3.96 (s, 3 H). MS (ESI):
calculated for Ci8FIgN203, 326.1, found 349.0
[M + Na]4, 324.9 [M - Hr. HPLC2: tR 14.65
min, purity > 99%.
(2-(2-(tiifluoromethyl)pheny1)-1H-imidazol-4- Yield: 83.8%; mp 75 - 77 C. IH
NMR (500
yl)(3,4,5-trimethoxypheny1)methanone (121a) MHz, CDC13) 5 10.37 (br, 1 H),
8.00-8.02 (m, 1
I-1), 7.87 (s, 1 H), 7.82-7.85 (m, 1 H), 7.69-7.74
(m, 1 H), 7_62-7.66 (m, 1 H), 7.25 (s, 2 H), 3.99
(s, 3 H), 3.98 (s, 6 H). MS (ESI): calculated for
C2011oF3N204, 406.1, found 429.1 [M + Nar,
405.0 [M - Hr. HPLC2: tR 13.98 min, purity >
99%,
(4-fluorophenyl)(2-(2- Yield: 91.1%; mp 152- 154 C. 11-1 NMR
(500
(trifluoromethyl)pheny1)-1H-i midazol-4- MHz, CDC13) 5 8.12-8.14 (m, 2 H),
7.97 (d, 1=
yl)methanone (121b) 7.5 Hz, 1 H), 7.82-7.85 (m, 2 H), 7.69 (t,
J = 7.5
Hz, 1 H), 7.61 (t, J = 8.0 Hz, 1 H), 7.22 (t, J =
9.0 Hz, 2 H). MS (BSI): calculated for
CoHmE4N20, 334.1, found 357.1 (M + Nar.
332.9 [M -H]. HPLC2: tR 15.10 min, purity >
99%.
(2-(4-(benzy1oxy)pheny1)-1H-imidazol-4- Yield: 16.5%; mp 191 - 193 T. 'H
NMR (500-
yl)(3,4,5-trimethoxyphenyl)methanone (12ja) MHz, CDC13) 5 10.22 (br, 1 H),
7.93 (d, J = 9.0
Hz, 2 H), 7.81 (s, 1 H), 7.37-7.47 (m, 5 H), 7.24
(s, 2 H), 7.11 (d,J= 8.5 Hz,2H), 5.16 (s, 2 H),
3.97 (s, 3 El), 3.95 (s, 6 H). MS (ESI): calculated
for C26H241µ1205, 444.2, found 467.1 [M + Nar,
442.9 [M - H]. HPLC2: tR 17.36 min, purity .
95.5%.
171
CA 3030689 2019-01-18
(2-(4-(benzyloxy)pheny1)-1H-i rnidazol-4- Yield:
84.7%; mp 212 - 214 C. 'H NMR (300
yl)(4-fluorophenyl)methanone (12jb) MHz, CDC13)
5 10.28 (hr. 1 H), 799-8.04 (m, 2
H), 7.92-7.95 (m, 2 H), 7.76 (d, J = 1.5 Hz, 1
H), 7.38-7.48 (m, 5 H), 7.20-7.25 (m, 2 H),
7.09-7.12 (in, 2 H), 5.16 (s, 2 H). MS (ES!):
calculated for C23H0PN202, 372.1, found 395.1
+ Na]. HPLC2: tR 17.97 min,,purity 97.8%.
(2-(4-hydroxyphen y1)- H-i mi dazol -4- Yield:
72.3%. mp 191-193 C. 1H NMR (500
yl)(3,4,5-trimethoxyphenyl)methanone (12ka) MHz, CD30D) 8 8.31 (s, 1 H), 7.90
(d, J = 8.5
Hz, 2 H), 7.31 (s, 2 H), 7Ø5 (s, 2 El), 3.95 (s, 6
H), 3.88 (s, 3 H). MS (ES!): calculated for
CI9F118N205, 354.1, found 355.1 IM +
352.9 [M - H]. HPLC2: tR 12.25 min, purity
98.7%.
(2-(4-(hydroxypheny1)-1H-irnidazol-4-y1)(4- Yield:
89.0%; mp 276 - 278 C. 11-1 NMR (500
fluorophenyl)methanone (12kb) MHz, CDC13)
5 8.31 (s, 1 H), 8.13 (q, J = 5.5
Hz, 3.0 Hz, 2 H), 7.93 (d, J= 8.5 Hz, 2 H), 7.38
J = 8.5 Hz, 2 H), 7.07 (d, I = 8.5 Hz, 2 H).
MS (ES!): calculated for Ci6HIIEN202, 282.1,
found 283.0 [M + H], 280.9 [M - HI. HPLC2:
tR 13.46 min, purity 97.65%.
(2-(4-bromopheny1)-1H-imidazol-4-y1)(3,4,5- Yield: 25.6%; mp 190-192 C. 111
NMR (500
trimethoxyphenyl)methanone (121a) MHz, CDC13) 5 7.99 (d, J = 8.5 Hz, 2
H), 7.92
(s, 1 H), 7.70 (d, J = 8.5 Hz, 2 H), 7.32 (s. 2
H), 4.03 (s, 3 H), 4.00 (s, 6 H). MS (ES!) calcd
for CIRHry RrN204 416.0, found 417.0 [M -4-
H]. HPLC2: tR 4.24 min, purity 98.8%.
(2-(4-(trifluoromethyDpheny1)-1H-imidazol-4- Yield: 85.3%; mp 195 - 196 C. 11-
1 NMR (500
yl)(3,4,5-tiimethoxyphenyl)methanone (12pa) MHz, CDC13) 5 8.22 (d, = 8.5 Hz, 2
H), 7.96
(s, 1 H), 7.83 (d, I = 8.5 Hz, 2 H), 7.34 (s, 2 H),
4.04 (s, 3 H), 4.00 (s, 6 H). MS (PSI) calcd for
C201417F3N204 406.1: found 407.1 [M + Hr,
HPLC2: tct 18.00 min, purity >99%.
(2-phenyl -1H-i midazol-1-y1)(3,4,5- Yield:
39.8%; mp 113 - 115 C. 111 NMR (500
trimethoxypheny Dmethanone (12aaa) MHz, CDC13)
5 7.53 (q, J = 5.0 Hz, 3.0 Hz., 2
H), 7.41 (d, J = 1.0 1-1z, I H), 7.33-7.35 (in, 3
H), 7.23 (d, J = 1.0 Hz, 1 H), 7.03 (s, 2 H), 3.93
(s, 3 H), 3.85 (s, 6 H). MS (ES!): calculated for
Cf911181µ1204, 338.1, found 339.1 [M +Hr.
HPLC2: tR 13.8 mm, purity 95.6%.
(4-methoxyphenyl)(2-phenyl-1H-imidazol-1- Yield:
56.3%; mp 68 - 70 C. 'H NMR (500
yl)methanone (12aba) MHz, CDCI3)
8 7.78 (d, I = 9.0 Hz, 2 H), 7.54-
7.56 (m, 2 H), 7.32-7.34 (m, 4 II), 7.21 (d, I =
1.0 Hz, 1 H), 6.93 (d, J 8.5 Hz, 2 H), 3.90 (s, 3
H). MS (ES!): calculated for Ci7H14N202, 278.1,
found 301.0 [M +Nar, 276.8 [M - Hf. HPLC2:
tR 14.72 min, purity 95.7%.
=
172
CA 3030689 2019-01-18
(4-fluoropheny1)(2-(p-toly1)-1H-imidazol-4- Yield: 95%; mp 115 - 117 C. 'H
NMR (500
yl)methanone Ha salt (12db-HCI) MHz, DMSO-d6) 5 8.20-8.23 (in, 2 H), 8.18
(s,
1 H), 8.04 (d, J= 6.5 Hz, 2 H), 7.42 (t, J = 8.0
Hz, 2 H), 7.37 (d, J= 7.0 Hz, 2 H), 2.38 (s, 3
H). MS (ESI): calculated for C171-114FCIN20,
316.1, found 281.0 [M - HC1 + H]t HPLC2: tR
17.16 min, purity >99%.
(4-fluorophenyl)(2-(4-methoxypheny1)-1- Yield: 90.2%; mp 148-150 C. 111
NMR (500
methyl-I H-imidazol-4-yl)methanone (12cba) MHz, CDC13) 5 8.45 (q, J= 8.5
Hz, 5.5 Hz, 2
H), 7.79 (s, 1 H), 7.63 (d, J= 8.5 Hz, 2 H),
7.16 (t,J=8.5 Hz, 2 H), 7.03 (d, J= 9.0 Hz, 2
H), 3.89 (s, 3 H), 3.82 (s, 3 H). MS (ESI) calcd
for C181-115 FN202 310.1, found 311.0 [M +
H]'. HPLC2: tR 4.01 min, purity 97.6%.
(1-benzy1-2-(p-toly1)-1H-imidazol-4-y1)(3,4,5- Yield: 92.8%; mp 135-137 C. 'H
NMR (500
trimethoxyphenypmethanone (12daa) MHz, CDCI3) 8 7.81 (s, 1 H), 7.80(d, J=
6.5
Hz, 2 H), 7.58 (d, I= 8.0 Hz, 2 H), 7.41-7.45
(m, 3 H), 7.31-7.33 (in, 2 H), 7.20 (d, J = 7.0
Hz, 2 H), 5.33 (s, 2 H), 3.99 (s, 3 H), 3.98 (s, 6
H), 2.47 (s, 3 H). MS (ESI) calcd for
C27H2c,N204 442.2, found 443.1 [M +
HPLC I: tR 4.28 min, purity > 99%.
(1-methyl-2-(p-toly1)-1H-imidazol-4-y1)(3,4,5- Yield: 87.4%; mp 110-112 'C. IH
NMR (500
=
trimethoxyphenyl)methanone (12dab) MHz, CDC13) 5 7.87 (s, 2 H), 7.86 (d, J
= 8.0
Hz, 1 H). 7.65 (d, I = 10 Hz, 2 H), 7.37 (d. J
Hz, 2 II), 4.01 (s, 6 H), 4.00 (s, 3 II), 3.90
(s, 3 H). MS (ESI) calcd for C211122N204
366.2, found 367.2 [M + Hr. HPLC1: tR 4.23
min, purity > 99%.
(2-(4-(di methyl ami no)pheny1)-1-((4- Yield: 34.1%; mp 147-149 C. 111 NMR
methox yphenyl)sulfony1)-1H-irnidazol-4- (CDC13, 500 MHz) 8 8.07 (q, J =
8.5 Hz, 5.5
yl)(4-fluorophenyl)methanone (12gba) Hz, 2 H), 7.78 (d, J= 9.0 Hz, 2 H),
7.41 (d, J=
8.5 Hz, 2 H), 7.39 (s, 1 H), 7.23 (t, J = 8.5 Hz, 2
H), 6.91 (d, J= 9.0 Hz, 2 H), 6.68 (d, J = 9.0
Hz, 2 H), 3.89 (s, 3 H), 3.08 (s, 3 H). MS (ESI)
calcd for C25H22FN304S 479.1, found 502.1 [M
+ Nar. HPLC2: tR 18.6 min, puritT 96.9%.
(3,4,5-trihydroxyphenyl)(2-(3,4,5- Yield: 66.1 %. mp 294 - 296 "C. H NMR
(500
tri hydroxypheny1)-1H-imidazol -4- MHz, CD30D) 5 8.07 (s, 1 H), 7.07 (s, 2
H),
ypmethanone (13ea) 7.02 (s, 2 H). MS (ESL): calculated for
C161-112N207, 344.1, found 345.01M + Hi, 342.9
[M - Hr. HPLC2: tR 3.62 min, pur1ty97.9%.
173
=
CA 3030689 2019-01-18
(2-(4-chloropheny1)-1H-imidazol-4-y1)(3,4,5- Yield: 79.3%; mp > 300 T. 'H
NMR (500
trihydroxyphenypmethanone (131a) MHz, CD30D) ö 8.02 (d, J = 8.5 Hz, 2 14),
737
(s, 1 H), 7.54 (d, J = 8.5 Hz, 2 H), 7.14 (s, 2 H).
MS (ES]): calculated for C161-1110N204, 330.0,
found 331.1 [1v1 + Na], 328.9 [M -
HPLC2: tR 11.9 rnM, purity 95.6%.
(2-(3,4-dihydroxypheny1)-1H-imidazol-4- Yield: 62.2 To; mp > 300 C. 'H NMR
(500
yl)(3,4,5-trihydroxyphenyl)methanone (13ha) MHz, CD30D) 8 8.11 (s, 1 H), 7.46
(d, J = 2.0
Hz, 1 H), 7.42 (dd, J = 8.5 Hz, 2.0 Hz, 1 H),
7.10 (s, 2 H), 7.02 (d, J = 8.5 Hz, 1 H). MS
(ESI): calculated for Ci6H12N206, 328.1, found
329.0 EM + flr, 326.9 [M - HT. HPLC2: tit
3.64 min, purity 97.9%.
2-(4-nitropheny1)-4,5-dihydro-IH-imidazole Yield: 70.3 %. 'H NMR (500 MHz,
CDC13)
(14x) 8.30 (d, J = 9.0 Hz, 2 H), 7.98 (d, J =
8.5 Hz, 2
1-1), 3.88-3.95 (m, 4 H). MS (EST): calculated for
C9H9N302, 191.10, found 191.9 [M + Hr, 189.7
-
2-(4-fluoropheny1)-4,5-dihydro-1H-imidazole Yield: 60.2 %. 'H NMR (500 MHz,
CDCI3)
(14b) 7.80 (q, J= 7.0 Hz, 2 H), 7.11 (d, J= 10.0
Hz, 2
11), 3.82 (br, 4 H). MS (MI): calculated for
C91-19FN2, 164.10, found 165 [NI + Hr.
2-(4-methoxypheny1)-4,5-dihydro-IH- Yield: 56.9 %. 11-1 NMR (500 MHz,
CDC13) 8
imidazole (14c) 7.84 (d, J = 8.5 Hz, 2 H), 6.94 (d, J =
9.0 Hz, 2
H), 3.87 (s, 3 H), 3.85 (br, 4 1-1). MS (ES1):
calculated for Ci0H12N20, 176.10, found 177.0
[M + Hr.
EXAMPLE 6
SYNTHESIS OF SELECTED INDOLYL-BENZOYL-IMIDAZOLE COMPOUNDS
[00392] The synthesis of 15xaa is outlined in Figure 12. This route was
originally
designed for the synthesis of 12xa, but the nonselectivity of the benzoylation
at the indole-2 and
imidazole-4 positions resulted in the formation of 15xaa, which is a closely
related but bulkier
analog of llxaa. The indole-5-carboxaldehyde 8x was protected by a
phenylsulfonyl group on
the indole NH to afford intermediate 8xa. 8xa was reacted with glyoxal and
ammonium
hydroxide to generate the 2-aryl-imidazole 9xa. Protection of the irnidazole
NH with
phenylsulfonyl gave the intermediate 10xaa which was coupled with 3,4,5-
trimethoxybenzoyl
chloride to produce 16xaa. Removal of the protecting group from 16xaa provided
15xaa.
100393] Synthesis of 1-(Phenylsulfony1)-1H-indole-5-carbaldehyde (8xa).
To a solution
of indole-3-carboxaldehyde (100 mmol) in ethanol (500 mL) at room temperature
was added
174
CA 3030689 2019-01-18
potassium hydroxide (110 equiv), the mixture was stirred until total
solubilization. The ethanol
was completely removed in vacuum and acetone (250 mL) added followed by
benzenesulfonyl
chloride (110 equiv). The precipitate was filtered off and the filtrate was
concentrated and
recrystallized from methanol to give a white solid. Yield: 32.6% 111 NMR (500
MHz, CDC13) 8
10.17 (s, 1 H), 8_25 - 8.39 (m, 2 H), 7.97 - 8.09 (m, 3 H), 7.69 (t, J = 7.33
Hz, 1 H), 7.59 (t, J =
7.5 Hz, 2 H), 7.39 - 7.54 (m, 2 H). MS (ES!) calcd for C151-1111103S 285.1,
found 286.0 [M + Hr.
[00394] Synthesis of (5-(4-(3,4,5-Trimethoxybenzoyl)-1112imidazol-2-y1)-
1H-indol-2-
y1)(3,4,5-trimethoxyphenyl)methanone (15xaa): To a solution of (1-
(phenylsulfony1)-2-(1-
(phenylsulfony1)-2-(3,4,5-trimethoxybenzoy1)-1H-indo1-5-y1)-1H-imidazol-4-
y1)(3,4 ,5-
trimethoxyphenyl)methanone (16xaa) (1 mmol) in ethanol (20 mL) was added
sodium hydroxide
(10 equiv) and stirred overnight in darkness. The reaction mixture was diluted
by 50 mL of water
and extracted by ethyl acetate (250 mL). The organic layer was dried over
magnesium sulfate and
concentrated. The residue was purified by flash column chromatography (hexane:
ethyl acetate
3:1) or recrystallized from water and methanol to give a white solid. Yield:
30-95%.
[00395] 5-(1H-Itnidazo1-2-y1)-1-(phenylsuffony1)-1H-indole (9xa). Yield:
12.0%. 111
NMR (500 MHz, DMSO-d6) 5 8.33 (d, J = 2.9 Hz, 2 H), 8.13 (d, J = 7.8 Hz, 2
14), 7.98 - 8.04 (m,
1 H), 7.62 - 7.67 (m, 1 H), 7.55 (d, J = 7.82 Hz, 2 H), 7.22 - 7.34 (m, 4 H).
MS (ES!) calcd for
C171113N302S 323.1, found 324,0 [M + Hr.
[00396] 1-(Phenylsulfonyl)-5-(1-(phenylsulfonyl)-1H-nnidazol-2-y1)-1H-
indole
(10xaa). Yield: 23.6%. 'I-1 NMR (500 MHz, CDCI3) 5 8.01 (d, J = 8.5 Hz, 1 H),
7.95 (d, J = 7.5
Hz, 2 H), 7.73 (d, J = 1.0 Hz, 1 1-1), 7.70 (d, J = 4.0 Hz, 1 H), 7.63-7.66
(m, 2 H), 7.52-7.56 (m, 3
H), 7.31-7.34 (m, 3 H), 7.22 (t, J = 8.5 Hz, 2 H), 7.17 (s, 1 H), 6.14 (d, J =
3.5 Hz, 1 H). MS
(ES!) calcd for C23Hr7N304S2 463.1, found 464.0 [M + Hr.
[00397] (1-(PhenylsulfonyI)-2-(1-(phenylsulfony1)-2-(3,4,5-
trimethoxybenzoy1)-1H-
indo1-5-y1)-1H-imidazol-4-y1)(3,4,5-trimethoxyphenyOmethanone (16xaa). Yield:
15.9%. /1-I
NMR (500 MHz, CDC13) 5 8.18 - 8.25 (m, 3 H), 8.04 (d, J = 8.1 Hz, 2H), 7.70-
7.78 (m, 2 H),
7.61 - 7.69 (m, 3 H), 7.55 (1,) 7.7 Hz, 3 H), 7.50 (s, 1 H), 7.38 (s, 2 H),
7.34 (s, 2 H), 6.94 (s, 1
H), 3.99 - 4.06 (m, 12 H), 3.94 - 3.99 (m, 6 H). MS (ESI) calcd for
C43H37N3012S2 851.2, found
852.1 [M + H]t.
175
CA 3030689 2019-01-18
[00398] (544-(3,4,5-Trimethoxybenzoy1)-1H-imidazol-2-y1)-1H-indol-2-
y1)(3,4,5-
trimethoxyphenypmethanone (15xaa). Yield: 45.9%; mp 239-241 C. ill NMR (500
MHz,
CDC13) ö 10.45 (s, I H), 9.44 (s, 1 H), 8.41 (s, 1 H), 8.04 (d, J = 8.5 Hz, 1
H), 7.86 (s, 1 H), 7.61
(d, J = 8.5 Hz, 1 H), 7.29 (s, 2 H), 7.26 (s, 2 H), 3.99 (s, 3 H), 3.95-3.97
(m, 15H). MS (ESI)
calcd for C311129N308 571.2, found 572.2 [M + H]. HPLC2: tR 4.09 min, purity
96.3%.
EXAMPLE 7
SYNTHESIS OF (2-(1H-INDOL-3-YL)-1H-IIVHDAZOL-4-YL)(3,4,5-
TRIMETHOXYPHENYL)METHANONE (17va) (FIGURE 13)
HN OMe
HN
OMe
OMe (17ya)
[00399] Synthesis of 1-(pheny1sulfony1)-1H-indole-3-carboxaldehyde
(8ya): To a
solution of indole 3-carboxaldehyde (8y) (100 mmol) in ethanol (500 mL) at RT
was added
potassium hydroxide (1.1 equiv). The mixture was stirred until total
solubilization. The ethanol
was completely removed in vacuum and the residual was dissolved in acetone
(250 mL) followed
by adding benzenesulfonyl chloride (1.1 equiv, 110 mmol). The reaction mixture
was stirred for
half hour. The precipitate was filtered off and the filtrate was concentrated
and recrystallized
from methanol to give a white solid. Yield: 33%. Ili NMR (500 MHz, CDC13) 5
10.17 (s, 1 H),
8,25-8.39 (m, 2 H), 7.97-8.09 (m, 3 H), 7.69 (t, J = 7.33 Hz, 1 H), 7.59 (t, J
= 7.5 Hz, 2 H), 7.39 -
7.54 (m, 2 H). MS (ESI) calcd for Ci5HIINO3S 285.1, found 286.0 [M + H].
[00400] Synthesis of 3-(111-imidazol-2-y1)-1-(phenylsulfony1)-1H-indole
(9ya): To a
solution of 1-(phenylsulfony1)-1H-indole-3-carboxaldehyde (8ya) (100 mmol) in
ethanol (400
mL) at 0 C was added a solution of 40% oxalaldehyde (glyoxal) in water (1.1
equiv, 110 mmol)
and a solution of 29% ammonium hydroxide in water (10 equiv, 1000 mmol). After
stirring for 2-
3 days at RT, the reaction mixture was quenched by water and extracted by
dichloromethane. The
organic layer was removed by vacuum and the residue was subjected to flash
column
chromatography with hexane/ethyl acetate (4:1-2:1) as eluent to yield the
titled compound as a
176
CA 3030689 2019-01-18
yellow powder. Yield: 12%.11-1 NMR (500 MHz, DMSO-d6) 5 8.33 (d, J = 2.9 Hz, 2
H), 8.13 (d,
J= 7.8 Hz, 2 H), 7.98 - 8.04 (m, 1 H), 7.62-7.67 (m, 1 H), 7.55 (d, J= 7.82
Hz, 2 H), 7,22 - 7.34
(m, 4 H). MS (ES!) calcd for C17Hi3N302S 323.1, found 324.0 [M + H].
11004011
Synthesis of 1-(phenylsulfony1)-3-(1-(phenyisulfony1)-1H-imidazol-2-y1)-1H-
indole (10ya): To a solution of 3-(1H-imidazol-2-y1)-1-(phenylsulfony1)-1H-
indole (9ya) (20
mmol) in anhydrous THF (300 mL) at 0 C was added sodium hydride (60%
dispersion in
mineral oil, 1.2 equiv, 24 mmol) and stirred for 20 min. Benzenesulfonyl
chloride (1.2 equiv, 24
mmol) was added and the reaction mixture was stirred overnight. After dilution
by 200 mL of
saturated NaHCO3 solution (aqueous), the reaction mixture was extracted by
ethyl acetate (600
mL). The organic layer was dried over magnesium sulfate and concentrated. The
residue was
purified by flash column chromatography (hexane: ethyl acetate 5:1) to give a
white solid. Yield:
40%. 114 NMR (CDC13, 300 MHz) 5 8.02-8.08 (m, 4 H), 7.72 (d, J = 1 . 5 Hz, 1
H), 7.35-7.60 (m,
8 H), 7.23 (d, J = 1.5 Hz, 1 H), 7.10-7.16 (m, 3 H). MS (ESI) calcd for
C23H17N304S2 463.1,
found 486.0 [M + Na]+.
[00402] Synthesis of (1-(phenylsulfony1)-2-(1-(phenylsulfony1)-1H-indol-3-
y1)-1 H -
imidazol-4-y1)(3,4,5-trimethoxyphenyl)methanone (17yaa): 'To a solution of 1-
(phenyl sul fony1)-3-(1-(phenylsulfony1)-1H-imidazol-2-y1)-1H-indole (10ya)
(5.0 mmol) in
anhydrous THF (100 mL) at -78 C was added 1.7 M tert-butyllithium in pentane
(1.2 equiv, 6.0
mmol) and stirred for 10 min. A solution of 3,4,5-trimethoxybenzoyl chloride
(1.2 equiv, 6.0
mmol) in THF was added at -78 C and stirred overnight. The reaction- mixture
was quenched
with 100 mL of saturated NaHCO3 solution (aqueous) and extracted by ethyl
acetate (300 mL).
The organic layer was dried over magnesium sulfate and concentrated. The
residue was purified
by flash column chromatography (hexane: ethyl acetate 3:1) to give a white
solid. Yield: 30%. 1H
NMR (500 MHz, CDCI3) 5 8.09 (d, J = 10 Hz, 1 H), 8.04 (d, J = 10 Hz, 2 H),
7.91 (s, I H), 7.76
(d, J = 5 Hz, 2 H), 7.65 (t, J = 10 Hz, 1 H), 7.55-7.58 (m, 5 H), 7.40 (s, 2
H), 7.33-7.36 (m, 3 H),
7.25 (t, J = 10 Hz, 1 H),4.05 (s, 3 H), 4.03 (s, 6 H). MS (ES!) calcd for
C33H27N308 657.0, found
680.1 [M + Nar
[00403] Synthesis of (2-(1H-
indol-3-y1)-1H-imidazol-4-y1)(3,4,5-
trimethoxyphenyl)methanone (17ya): To a
solution of ( I -(phenylsulfony1)-2-(1-
(phenylsulfony1)-1H-indo1-3-y1)-1H-imidazol-4-y1)(3,4,5-
trimethoxyphenyl)methanone (17yaa)
(1 mmol) in ethanol (40 mL) and water (4 mL) was added sodium hydroxide (10
equiv, 10 mmol)
177
CA 3030689 2019-01-18
and stirred overnight under refluxing condition in darkness. The reaction
mixture was diluted by
50 nil. of water and extracted by ethyl acetate (200 mL). The organic layer
was dried over
magnesium sulfate and concentrated. The residue was purified by flash column
chromatography
(hexane: ethyl acetate 1:1) to give a yellow solid. Yield: 60%. NMR (500
MHz, CD30D)
8.31 (d, J 6.5 Hz, 1 H), 7.99 (s, 1 H), 7.90 (s, I H), 7.48-7.52 (m, 3 H),
7.24-7.28 (m, 2 H), 4.00
(s, 6 H), 3.93 (s, 3 H). MS (ES!) calcd for C21Hi9N304 377.1, found 400.1 [M +
Mp 208-
210 T.
EXAMPLE 8
SYNTHESIS OF (2-(1H-INDOL-5-YLAMINO)THIAZOL-4-YL)(3,4,5-
TREVIETHOXYPHENYL)METHANONE (COMPOUND 55) (FIGURE 15).
[00404] A
mixture of 5-nitro-1H-indole (11 g, 67.9 mmol) and Pd/C (5%; 1 g), dissolved
in ethanol (50 mL), was hydrogenated for 3 h at 40 psi. The reaction mixture
was filtered and the
excess of ethanol was evaporated under reduced pressure. Solid product was
recrystallized from
hexane to obtain the pure compound 5-aminoindole (55-1). Yield: 92.5%. NMR
(500 MHz,
CDCI3): 8 7.96 (br, 1 H), 7.20 (d, 1 H), 7.13 (s, 1 H), 6.95 (s, 1 H), 6.67
(dd, 1 H), 6.37 (s, 1 H),
3.50 (s, 2 H). MS (ES!) rri/z 133.0 (M + H)".
[00405) A
solution of 5-aminoindole (8 g, 60.6 mmol) in acetone (150 mL) was reacted
with benzoylisothiocyanate (9.88 g, 60. mmol) at RT for about 4h. The
resulting solid was
filtered and treated with 2 N NaOH in THF (120 mL). The mixture was refluxed
for about 6 h
and allowed to warm to RT. The solvent was evaporated off under vacuum. The
residue was
diluted with water (20 mL) and neutralized to pH 7 with 1 N HCI. The resulting
solid was filtered
and dried under vacuum to afford 5-indolylthiourea (55-2). 5-Indoly1 thiourea
(0.01 mot) and
ethyl bromopyruvate (0.011 mol) were dissolved in 3 mL ethanol and held at
reflux for 2 h. The
= reaction was cooled, the crystalline ethyl 2-(111-indo1-5-
ylamino)thiazole-4-carboxylate (55-3)
was collected by filtration and washed with ethanol. Refluxing the mixture of
ethyl esters with
the NaOH-ethanol solution gave 2-(1H-indo1-5-ylamino)thiazole-4-carboxylic
acid (55-4) which
was used directly in next steps: To a mixture of the crude acid (2.5 mmol),
EDCI (2.9 mmol),
HOBt (2.6 mmol) and NMM (5.3 mmol) in CH2Cl2 (30 mL) was added HNCH30CH3HCI
salt
(2.6 mmol) and stirring continued at RT for overnight. The reaction mixture
was diluted with
CH2Cl2 (20 mL) and sequentially washed with water, satd. NaHCO3, brine and
dried over
178
CA 3030689 2019-01-18
MgSO4. The solvent was removed under reduced pressure to yield a crude
product, which was
purified by column chromatography to obtain pure compound 2-(1H-indo1-5-
ylamino)-N-
methoxy-N-methylthiazole-4-carboxamide (55-5) (45.6% yield for overall 5
steps). At -78 C, to
a solution of 5-bromo-1,2,3-trimethoxybenzene (1.235 g, 5.0 mmol) in 30 mL THF
was charged
n-BuLi in hexane (2.5 N, 2.4 mL, 6 mmol) under Ar2 protection and stirred for
10 min Weinreb
amide (1 mmol) in 10 mL THF was added to lithium reagent and allowed to stir
at RT for 2 h.
The reaction mixture was quenched with satd. NI-WI, extracted with ethyl
ether, dried with
MgSO4. The solvent was removed under reduced pressure to yield a crude
product, which was
purified by column chromatography to obtain pure compound 55 (51.7% yield). 'H
NMR (300
MHz, CDCI3) 8 8.29 (br, 1 II), 7.68 (d, 1 H), 7.46 (s, 2 H), 7.39 (s, 1 11),
7.36 (s, 1 H),7.28 ¨ 7.26
(m, 1 H), 7.15-7.12 (m, 1 H), 6.55 (m, 1 H), 3.93 (s, 3 H), 3.89 (s, 6 H). MS
(ESI) nik 432.1 (M
+ Na), 408.0 (M - H)",
EXAMPLE 9
SYNTHESIS OF OUINOLINE- AND ISOOUINOLINE-ARYL COMPOUNDS (FIGURE
A series of compounds were prepared by Suzuki coupling of 7-bromo-1-
chloroisoquinoline with
various arylboronic acids.
=
Synthesis of 1-Chloro-7-(1H-indo1-5-y1)-isoquinoline (6d) (Figure 16C):
B(01-1)2
Pd{PP113)., K2CO3,,
Br 414r. DMF, H20, reflux 5h
CI NH CI
6d
[00406] A mixture of 7-bromo-1-chloroisoquinoline (0.50 g, 2.1 mmol), 5-
indoleboronic
acid (0.40 g, 2.5 mmol), tetrakis(triphenylphosphene)palladium (0.035 g, 0.08
mmol), potassium
carbonate (2.1 mL, 2 M, 4.1 mmol), N,N-dimethylformamide (11 mL) was stirred
while purging
the headspace with argon for 30 min. The mixture was then brought to reflux
for 16 h before
being allowed to cool to RT. The mixture was filtered through a bed of silica
gel, diluted with
water (50 mL), and extracted with ethyl acetate (50 mL). The organic layer was
separated and
washed with NaOH (2 x 20 mL, 10 % aq.), water (5 x 30 mL, until refractive
changes were no
longer seen at the organic-aqueous interface), and ammonium chloride (20 mL,
sat.). The organic
179
CA 3030689 2019-01-18
layer was then adsorbed onto silica gel and flash-chromatographed (ethyl
acetate/hexanes) to
afford 0.14 g (25 %) of a yellow solid. MS (ESI): calculated for C171111CIN2,
278.1, found 301.0
EM + Na]. NMR (300 MHz, DMSO-d6) 5 6.56 - 6.58 (m, 1 H), 7.44 (t, J =
2.77 Hz, 1 H),
7.57 - 7.59 (m, 2 H), 7.93(m, 1 H), 8.04 (s, 1 H), 8.13 - 8.20 (in, 1 H), 8.27
- 8.31 (m, 2 H), 8.43
(m, 1 H), 11.25 (brs, 1 H).
1,7-Bis-(1H-indo1-5-y1)-isoquinoline (6b) (Figure 16E):
OH
t
/ OH
N "gr
-N
Br
Cl Pd(PPII3)4. K2CO3,
DMF. H2O. reflux 5-18 h 6b \ NH
100407] A mixture of 7-bromo-1-chloroisoquinoline (0.20 g, 2.1 mmol), 5-
indoleboronic
acid (0.80 g, 5.0 mmol), tetrakis(triphenylphosphene)palladium (0.19 g, 0.16
mmol), potassium
carbonate (2.1 mL, 2 M, 4.1 mmol), N,N-dimethylformamide (11 mL) was stirred
while purging
the headspace with argon for 30 min. The mixture was then brought to reflux
for 16 h before
being allowed to cool to RT. The mixture was filtered through a bed of silica
gel, diluted with
water (50 mL), and extracted with ethyl acetate (50 mL). The organic layer was
separated and
washed with NaOH (2 x 20 mL, 10 % aq.), water (5 x 30 mL, until refractive
changes were no
longer seen at the organic-aqueous interface), and ammonium chloride (20 mL,
sat.). The organic
layer was then adsorbed onto silica gel and flash-chromatographed (ethyl
acetatelhexanes) to
afford 0.29 g (39 %) of a yellow solid. MS (ESI): calculated for C251-117N3,
359.1, found 360.2 [M
+ Hr 382.1 [M + Nar, and 358.0 [M - H].IH NMR (500 MHz, DMSO-d6) 8 6.46 - 6.50
(n, 1
H) 6.52 - 6.59 (m, 1 H) 7.34 - 7.36 (m, 1 H) 7.36 - 7.41 (m, 2 H) 7.42 - 7.52
(m, 3 H) 7.58 (d,
J=8.30 Hz, 1 H) 7.81 (dd, J=5.49, 5.00 Hz, 2 H) 7.92 (s, 1 H) 8.08 - 8.17 (in,
2 H) 8.33 (s, 1 H)
8.54 (d, J=5.61 Hz,! H) 11.18 (br. s., 1 H) 11.30 (br. s., 1 H) ppm.
1-(4-Fluoro-phenyl)-7-(1H-indol-5-y1)-isoquinoline (6c) (Figure 16D):
180
CA 3030689 2019-01-18
=
B(oF02 ,N
pceph,)õ,K2e03i. N
,N
DMF, H20, reflux 5 h H =
CI
6C F
6d
[00408] A mixture of bd (0.20 g, 0.72 mmol), 4-fluorophenylboronic acid
(0.12 g, 0.86
mmol), tetralcis(triphenylphosphene)palladium (0.033 g, 0.03 mmol), potassium
carbonate (0.72
mL, 2 M, 1.4 mmol), N,N-dimethylformamide (22 mL) was stirred while purging
the headspace
with argon for 30 mm. The mixture was then brought to reflux for 16 h before
being allowed to
cool to RT. The mixture was filtered through a bed of silica gel, diluted with
water (50 mL), and
extracted with ethyl acetate (50 mL). The organic layer was separated and
washed with NaOH (2
x 20 mL, 10 % aq.), water (5 x 30 mL, until refractive changes were no longer
seen at the
organic-aqueous interface), and ammonium chloride (20 mL, sat.). The organic
layer was then
adsorbed onto silica gel and flash-chromatographed (ethyl acetate(hexanes) to
afford 0.038 g (16
%) of a yellow solid. MS (ES!): calculated for C23H15FN2, 338.12, found 339.2
[M + H]' and
361.2 [M + Na]. 11-1 NMR (300 MHz, DMSO-d6) 8 6.47 - 6.55 (m, 1 H), 6.80 (d, J
= 9.16 Hz, 2
H), 7.38 - 7.45 (m, 2 H), 7.47 - 7.62 (m, 3 H), 7.72 (d, J = 8.85 Hz, 2 H),
7.79 - 7.96 (m, 3 H),
11.18 (br. s., 1 H).
[00409] 1,7-Bis-(4-fluoro-phenyl)-isoquinoline (40) (Figure 16A).
,N
FO
F
[00410] MS (ES!): calculated for C211-113F2N, 317.10, found 318.1 [M +
Hr, 340.1 [M +
Na], and 315.9 [M - H]. IHNMR (500 MHz, DMSO-d6) 8 7.31 (br. s., 1 H) 7.31 -
7.37 (m, 2 H)
7.39 (br. s., 1 H) 7.41 (t, J = 8.54 Hz, 2 H) 7.72 - 7.77 (m, 2 H) 7.78 - 7.84
(m, 2 H) 7.89 (br. s., 1
H) 7.90- 7.99 (m, 1 H) 8.09- 8.19 (m, 3 H) 8.59 (br. S., 1 H) 8.60 - 8.65 (m,
1 H).
=
181
CA 3030689 2019-01-18
[00411] Synthesis of 7-Bromo-1-(4-fluoro-benzenesulfony1)-1,2,3,4-tetrahydro-
quinoline (43) and 1-(4-
Fluoro-benzenesulfony1)-7-(1H-indol-5-y1)-1,2,3,4-
tetrahydroquinoline (41). (Figure 16B).
1161
Ole _a_p. BrN b
Br 02S 023 ill
43 41 F
[00412] 7-
Bromo-1,2,3,4-tetrahydroquinoline (0.60 g, 2.8 mmol) was stirred with 4-
fluorophenylsulphonyi chloride (1.65 g, 8.49 mmol) in pyridine (5 mL) at 80 C
for 3 h. The
mixture was cooled, concentrated, and the residue was chromatagraphed
(Et0Ac/Hexanes on
SiO2) to give 845 mg of a brown solid (81 %) of compound 43. CI5H13BrFNO2S
368.98, found
394.0 [M + Na]' and 367.8 [M - NMR
(500 MHz, CDCI3) 8 1.58 - 1.67 (rn, 2 H) 2,41 (t,
J = 6.71 Hz, 2 H) 3.72 - 3.82(m, 2 H) 6.89 (d, J= 8.30 Hz, 1 H) 7.08- 7.17(m,
2 H) 7.18 -7.24
(m, 1 H) 7.59 - 7.68 (m, 2 H) 7.92 - 8.01 (m, 1 H).
[00413] 43 (0.46 g, 1.3 mmol), 5-indoleboronic acid (0.26 g, = 1.6 mmol),
tetrakis(triphenylphosphene)palladium (0.031 g, 0.03 mmol), potassium
carbonate (1.35 mL, 2-
M, 2.7 mmol), and N,N-dimethylformamide (135 mL) were stirred while purging
the headspace
with argon for 30 min. The mixture was then brought to reflux for 16 h before
being allowed to
cool to RT. The mixture was filtered through a bed of silica gel, diluted with
water (50 mL), and
extracted With ethyl acetate (50 mL). The organic layer was separated and
washed with NaOH (2
x 20 mL, 10 % aq.), water (5 x 30 mL, until refractive changes were no longer
seen at the
organic-aqueous interface), and ammonium chloride (20 mL, sat.). The organic
layer was then
adsorbed onto silica gel and flash-chromatographed (ethyl acetate/hexanes) to
afford 0.38 g (77
%) of a white crystalline solid of compound 41. MS (ES!): calculated for C231-
119FN202S, 406.12,
found 404.9 [M - Hy and 429.1 [M + Nal+. 11-1 NIVIR (500 MHz, DMSO-d6) 5 1.56 -
1.66 (m, 2
H)2,48 (t, J = 6.59 Hz, 2 H) 3.76 - 3.86 (m, 2 H) 6.46 - 6.56 (rn, 1 H) 7.14
(m, J = 7.81 Hz, 1 H)
7.33 - 7.37 (m, 1 H) 7.38 - 7.45 (in, 4 H) 7.49 (in, J = 8.54 Hz, 1 H) 7.66 -
7.74 (m, 2 H) 7.74 -
7.81 (m, 1 H) 7.85 - 7.94 (in, 1 H) 11.17 (br. s., 1 H).
182
CA 3030689 2019-01-18
[004141 7-Bromo-2-(4-fluoro-benzenesulfonyI)-1,2,3,4-tetrahydro-isoquinoline
(42)
(Figure 16B).
40 101 N, -0 3 Br b
NH
-0 --I.
42 * 44 s
[00415] Yield 23 %. Ci5Hi3BrFNO2S, 369.0, found 392.0 Uvl + Na}+ and
367.7 IM - Hr.
11-1 NMR (500 MHz, DMSO-d6) 5 2.75 - 2.82 (m, 2 H) 3.32 (t, J = 6.10 Hz, 2 H)
4.24 (s, 2 H)
7.07 (d, J = 8.30 Hz, 1 H) 7.29 -7.37 (m, 1 H) 7.37 - 7.43 (m, 1 H) 7.47 (t, J
= 8.79 Hz, 2 H) 7.87 .
- 7.93 (m, 2 H).
[00416] 2-(4-Fluoro-benzenesulfony1)-7-(111-indo1-5-y1)-1,2,3,4-
tetrahyd ro-
. isoquinoline (44).
[00417] Yield 77 %. 1H NMR (500 MHz, DMS0-4) 8 2.84- 2.91 (m, 2 H) 3.35 (t,
J=5.98
Hz, 2 H) 4.30 (s, 2 H) 6.44 -6.48 (m, I H) 7.17 (d, J = 7.81 Hz, 1 H) 7.32-
7.40 (m, 2 H) 7.41 -
7.51 (m, 3 H) 7.75 - 7.79 (m, I H) 7.89 - 7.96 (m, 1 H) 11.13 (br. s., I H).
EXAMPLE 10
WATER SOLUBILITY OF ARYL-BENZOYL-LMIDAZOLE (ABI) COMPOUNDS (Figure
in
[00418] Determination of water solubility. To determine water
solubility, 1 mg of each
compound was suspended in 1 mL water and shaken for 48 h at room temperature
(RT). The
suspension was centrifuged at 10,000 rpm for 10 min and filtered on 0.22 pm
filter.
Concentrations of each compound were measured by LC-MS, consisting of an HP
S1100 HPLC
instrument (Agilent, Foster ceity, CA) and a Bruker ESQUIRE MS detector with
electrospray/ion
trap instrument in positive ion mode (Bruker, Fremont, CA). For HPLC, a
reverse phase Nova-
palc C18 column (150mm x 3.9 mm, Waters, Milford, MA) was used. The mobile
phase was
composed of 20:80 v/v water/acetonitrile. For MASS, the peak was extracted at
382 m/z (for
imidazole compounds) and 399 rn/z (for thiazole compounds) respectively. The
concentration of =
each compound was calculated by MS peak area according to the following
calibration equation:
y=1.3295x + 114.24 (R2=1.00). To make the standard curve (Figure 17) from
which the the
equation was derived, 50, 100 L of each 100 g/mL, 10 i.tg/mL of ABI compound
12ga, and its
183
CA 3030689 2019-01-18
corresponding thiazole analog, as well as CA-4 (see Figure 19 for structure)
in acetonitrile, were
injected into HPLC and monitored by mass spectroscopy. The amount (ng) in each
injection was
plotted again its relative mass peak area to generate the standard curve in
Figure 17.
[00419] The HPLC retention times of ABI compound 12ga (1.5 min) was
compared to its
corresponding thiazole analog (2.2 min) using 80/20 methanol/water mobile
phase at 1 mUmin
flow rate and a reversed phase column, indicating that the imidazole
derivative was more
hydrophilic than its corresponding thiazole analog. The calculated logP values
for ABI compound
12ga and the corresponding thiazole analog were approximately 2.9 and 4.4,
respectively. The
water solubility of compound 12ga was 13 g,/mL, or about 200 times greater
than its thiazole
counterpart (72 ng,/mL).
=
EXAMPLE 11
BIOLOGICAL EVALUATION OF COMPOUNDS OF THIS INVENTION:
Example 11A: In Vitro Cell Growth Inhibitions.
[00420] Cell Culture and Cytotoxicity Assay of Prostate Cancer and
Melanoma. All
cell lines were obtained from ATCC (American Type Culture Collection,
Manassas, VA, USA),
while cell culture supplies were purchased from Cellgro Mediatech (Herndon,
VA, USA). We
examined the antiproliferative activity of our anti-tubulin compounds in four
human prostate
cancer cell lines (LNCaP, DU 145, PC-3, and PPC-1) and two human melanoma cell
lines (A375
and WM-164). Human ovarian cell line OVCAR-8 and its resistant cell line that
over-expresses
P-gp (NCUADR-RES) were used as MDR models. Both ovarian cell lines were
obtained from
National Cancer Institutes (NCI). All cell lines were tested and authenticated
by either ATCC or
NCI. All prostate cancer and ovarian cancer cell lines were cultured in RPM!
1640, supplemented
with 10% fetal bovine serum (FBS).
[00421] Melanoma cells were cultured in DMEM, supplemented with 5% FBS,
1%
antibiotic/antirnycotic mixture (Sigma-Aldrich, Inc., St. Louis, MO, USA) and
bovine insulin (5
p.g/mL; Sigma-Aldrich). The cytotoxic potential of the anti-tubulin compounds
was evaluated
using the sulforhodarnine B (SRB) assay after 96 h of treatment.
184
CA 3030689 2019-01-18
All of the reported compounds were first evaluated for cytotoxicity in the
mouse
melanoma cell line B16-F1, human melanoma cell lines (A375 and WM-164) and
prostate cancer
cell lines (DU145, PC-3, "LNCaP, PPC-1). Compound lh and ABT-751 (E7010,
Abbott
Laboratories/Eisai Co Ltd), which has entered phase II clinical studies in
treating patients with
different cancers, were included in the assays as examples of colchicine-site
binding agents. IC50
values for cell growth inhibition are shown in Tables 1, 2 and 3.
=
185
CA 3030689 2019-01-18
Results:
Table 1. SAR of B ring Optimizing Compounds
0-
.
alk = = .
IC50 SEM (Oil)
lb/ 0- \ Bring _____________________________________________
101 B 16-Fl A375 DU 145 PC-3 LNCaP PPC-1
la 1,3-phenyl 500 200 87 15 178 81 234 85
lb 4,6-pyrimidine >30000 >30000 6900 8300 7000 3700
lc 2,6-pyridine 39 I2 30 14 33 3 32 2 27 2 25 1
hl 2,5-furan 151 24 27 8 35 21 23 20
le 2,5-thiazole 12500 5200 13600 3800 >10000
>10000 >10000 >10000
If 2,4-thiophene 72 I5 15 6 26 12 17 15
lg 1,4-piperidine >30000 >30000 >20000 >20000
>20000 >20000
lb 2,4-thiazole 55 5 28 5 7114 211-1 28+4 4315
li 3,5-isoxazole >30000 >30000 >10000 >10000 >10000 >10000
36a 2,4-ox azo le 600 200 300 100 292 294 310
324
35a 2,4-oxazol Me 6500 800 500 100 1200 100 1200 100
1200 100 11001100
Table 2. SAR of Carbonyl Linker Optimizing Compounds
c=-=
-
1050 SEM (nM)
561 o- X lin ker
B16-F I A375 WM-164 DU 145 PC-3 LNCaP
PPC-1
Hi C=0 15515 28 5 64+4 71-14 2111 2814 43 5
2a C=CMe2 3800 1300 1900 800 3704 1200 2650 2470 1390
2040
2b CHOH >30000 >30000 ND >10000 ,>10000 >10000 >10000
2c-trans syn-C=C-CN 5400-12100 460111500 ,4900 1300 , 2280 890 580
900
2c-cis anti- C=C-CN 1200 300 1200 400 1030 200 -10000 , -10000
1990 -10000 ,
2d-cis syn- C=N-NH2 2000 800 ,900 300 ND 1210 1120 1800 872
2d-trans Anti- C=N-NH2 1800 700 600 200 ND 1210 1040 ,1300
;966
2e-cis syn- C= N-OH 300 100 200 1 00 ND* 102 ' 120 189
160
2e-trans anti- C.=N-OH 11400 2100 ,7800 1200 ND >10000 -10000 >10000
>10000
2f-cis syn- C=N-0Me 3800 1600 2900 1200 34001:1800 >10000 ,r>I0000
>10000 >10000
2f-trans 4nti-C=N-0Me >10000 >10000 >10000 >10000 >10000
>10000 >10000
¨
2g CONH >30000 >30000 ND ?10000 >10000 >10000
>10000
2h NI-ICO >30000 >30000 ND õ>10000 ,>10000
>10000 >10000
,
2i Bond (none) >10000 >10000 >10000 >10000 7.10000
>10000 >10000 _
,
2j C=N-CN 60 21 28 12 27 13 42 2 27 1 73 2 20 1
. - -
3a eis-C=C 11000 2800 46500 2330010600 5800 >10000 >10000
?10000 >10000
, .
3b trans-C=C 32800 1300(b10000 /30800 12000>10000 ,>10000 ,>10000 >10000
,
4a iS 2400 900 1600 400 2000 1200 , >10000 >10000 2300 200
2300 100 ,
46 SO2 >10000 >10000 ?10000 ,>10000 >10000 >10000
>10000
4c SC) ,
>10000 >10000 >10000 >10000 >10000 >10000
t>10000
4t1 SO5NH2 >10000 >10000 >10000 >10000 >10000
>10000 >10000
- - 5 *NT) = Not determined ¨
186
CA 3 0 3 0 6 8 9 2 0 1 9 -0 1 -1 8
Table 3. Antiproliferative Activity of Modified Compounds with Improved
Aqueous Solubility
0 _____________________________________________________________
1050 SEM (nM)
sr14 1-{,3:y Apart __________________________________________________
IP B16-F1 A375 DU 145 PC-3 LNCaP PPC-1
58a 4-0TB DMS Ph 500 200 700 300 434 30 183 24 , 549
246 8
- ______________________________________________________________________
21 4-0HPh 110 100 116 87 103 76
62a 2-indoly1 43 21 19 9 32 24 28 28
66a 5-indol yl 25 13 8 1 13 7 10 8
- ______________________________________________________________________
68a 4-BocNHCH2Ph 2900 400 7900 500 4400 3100 - 2600 2700
Zr '4-NH2CH2Ph ' 38 11 41 13 25 80 - 13 34
2s 4-NHMeCH2Ph >10000 >10000 -10000 >10000 114 80 -
1000
_______________________________________________________________________ _
2u 4-NMe2CH2Ph >10000 >10000 >10000 >10000 1025 200 >10000
5a PhNH 65 12 45 8 70 4 57 3 51 1 54 1
_
5Hb 4-CH3PhNH ND* ND 35 1 38 2 35 1 36 1
¨ ______________________________________________________________________
Sc 4-FININH ' ND ND 63 1 43 1 41 1 37 1
1 h Ph 55 5 28 5 71 4 21 1 ' 28+4 43 5
ABT-751 2127 351 - 1111 108 839 719 786 89
- 658 117 701 307
*ND_ Not determined
[00422] SAR of
alternative "B" ring molecules. The first series was targeted to
alternatives to the thiazole "B" ring. Accordingly, a series of heterocyclic
"B" rings were
examined. As shown in Table 1, the successful replacements of the thiazole
were pyridine lc,
furan id and thiophene if. The IC5os (12 nM - 35 nM against prostate cancer
cells) are close to
the thiazole compound lh. Introducing phenyl (la), oxazoline (35a), and
oxazole (36a)
maintained activity in the hundreds of nanomolar range. But introducing of
pyrirnidine (lb. IC5o:
3.7-8.3 M), a reversed 2,5-thiazole and 3,5-isoxazole (le and li, IC50: > 10
AM) caused obvious
losses of potency. Modification of "B" ring to the saturated ring of
piperidine (1g) also totally
abolished activity (1050>20 1.LM).
[00423] SAR of
Alternative Linkers. In vitro hepatic metabolic stability studies revealed
that the carbonyl linker between "B" and "C" rings in SMART compounds caused
short half lives
(5-17 min) primarily due to carbonyl reduction. For the sake of blocking this
ketone reduction to
the inactive hydroxyl linker compound 2b, the carbonyl linker in the second
series of compounds
187
CA 3030689 2 0 1 9 - 0 1 -1 8
was modified (Table 2). The carbonyl linker was replaced with double bonds
(2a, 3a and 3b),
amides (2g, 2h), oximes (2e-cis,trans and 2f-cis,trans), hydrazide (2d-cis, 2d-
trans),
acrylonitriles (2c-trans, 2c-cis), cyanoimine (2j), sulfonyl amide (4d),
sulfur ether (4a), sulfonyl
and sulfinyl compounds (4b, 4c). A direct link compound 21 without any linker
between "B" and
"C" rings was also prepared. Among these linker modifications, only cyanoimine
linkage (2j)
showed promising potential (20 ¨ 60 nIVI) compared with carbonyl compound lh,
but an in vitro
metabolism study showed that the half life of 2j in human liver rnicrosome was
less than 5 min.
This suggested that although the ketone reduction is blocked, it might
introduce a new metabolic
liability in compound 2j. The isomer pairs of compounds containing double
bonds, oxime's and
hydrazides were separated. Compound 3a was designed to mimic the structure of
CA-4, (Figure
19) which contain a cis-C=C between two aryl rings, unfortunately 3a and other
isomer pairs lost
activity after replacing the C=0 linker. One interesting phenomenon is syn-
isomer of 2e-cis (0.1
¨ 0.3 1.i.M) showed 10 fold more activity than its anti-isomer 2e-trans (>10
ilM). The half life of
2e-cis in human liver microsome is extended to 35 min, while half lives of
compounds 2d can be
prolonged to 55 min. But decreased activity (-11.1M) of 2d also reduced their
potency.
Example JIB: Aqueous Solubility of compounds of the invention.
[00424] The solubility of drugs .was determined by Multiscreen
Solubility Filter Plate
(Millipore Corporate, Billerica, MA) coupled with LC-MS/MS. Briefly, 198 1.11.
of phosphate
buffered saline (PBS) buffer (pH 7.4) was loaded into 96-well plate, and 2 tL
of 10 inM test
compounds (in DMSO) was dispensed and mixed with gentle shaking (200-300 rpm)
for 1.5 h at
RT (N = 3). The plate was centrifuged at 800g for 5 min, and the filtrate was
used to determine
its concentration and solubility of test compound by LC-MS/MS as described
below.
[00425] Introducing polar and ionizable groups into the anti-tubulin
agents. One
major limitation of the SMART agents is low aqueous solubility. Surfactants
such as Tween 80,
were used to study in vivo SMART behavior, accordingly favorable results were
obtained. But
these surfactants are biologically active and are responsible for many side
effects. In addition, it
was thought that low aqueous solubility of lb resulted in low oral
bioavailability (3.3%, Table
4). In the third series of compounds, the aqueous solubility was successfully
increased without
impacting the potency by introducing polar groups like hydroxyl and. indolyls.
In addition,
ionizable groups like amino and alkylamino groups were also introduced into
"A" ring para-
188
CA 3030689 2019-01-18
position. As shown in Figure 5 and Table 3, introducing indolyl groups to the
"A" ring
especially 5-indolyl (66a, 7 ¨ 25 nM) increased the potency compared with the
4-0H compound
21 (76-116 nM). Aminomethyl -CH2NH2 at the "A" ring para position also
maintained potency
(2r, 13-80 nM), but p-NHNIe (2s) or p-NMe2 (2u) abrogated activity. As shown
in Figure 18,
analytical measurement to estimate aqueous solubility showed that indolyl
compound 66a
increased solubility in PBS from 1.1 trg/mL (compound 1h) to 3.8 pg/mL.
Aminomethyl
compound 2r was converted to the HCI salt, which increased solubility over 35-
fold (>
35 g/mL). Although compound 2r showed satisfactory aqueous solubility, the
phannacokinetic
studies showed this compound still had very poor bioavailability (F% = 0.2%).
It was thought
that compound 2r was ionized in the stomach, and therefore not absorbed into
the circulation
system.
Example I IC: Phatmacokinetic studies
[00426] Pharmacokinetic Study. Female Sprague-Dawley rats (n = 3 or 4;
254 4 g)
were purchased from Harlan Inc. (Indianapolis, IN). Rat thoracic jugular vein
catheters were
* purchased from Braintree Scientific Inc. (Braintree, MA). On arrival at the
animal facility, the
animals were acclimated for 3 days in a temperature-controlled room (20-22 C)
with a 12 h
light/dark cycle before any treatment. Compound lb was administered
intravenously (i.v.) into
the jugular vein catheters at a dose of 2.5 mg/kg (in DMSO/PEG300, 2/8),
whereas 5Ha and 5Hc
were dosed at 5 mg,/kg (in DMSO/PEG300, 1/9). An equal volume 'of heparinized
saline was
injected to replace the removed blood, and blood samples (250 iaL) were
collected via the jugular
vein catheters at 10, 20, 30 min, and 1, 2, 4, 8, 12, 24 h. Compounds lh, 5Ha
and 5Hc were
given (p.o.) by oral gavage at 10 mg/kg (in Tween80/DMSO/1120, 2/1/7). All
blood samples (250
4) after oral administration were collected via the jugular vein catheters at
30, 60, 90 min, 120
min, 150 min, 180 min, 210 min, 240 min, and 8, 12, 24 h. Heparinized syringes
and vials were
prepared prior to blood collection. Plasma samples were prepared by
centrifuging the blood
samples at 8,000 g for 5 min. All plasma samples were stored immediately at -
80 C until
analyzed.
[00427] Analytes were extracted from 100 L of plasma with 200 I.tL of
acetonitrile
containing 200 nM the internal standard ((3,5-dimethoxyphenyl)(2-pheny1-1H-
imidazol-4-
yOmethanone). The samples were thoroughly mixed, centrifuged, and the organic
extract was
189
CA 3030689 2019-01-18
transferred to autosampler for LC-MS/MS analysis. Multiple reaction monitoring
(MRM) mode,
scanning nik 356 188 (compound 1h), miz 371 -+ 203 (compound 5Ha), ink 389
¨* 221 -
(compound 5Hc), and in/z 309 171 (the internal standard), was used to
obtain the most
sensitive signals. The pharmacokinetic parameters were determined using non-
compartmental
analysis (WinNonlin, Pharsight Corporation, Mountain View, CA)
Results:
Table 4. Pharmacokinetic Parameters for Compounds Tested in vivo.
lh 2r 5Ha 5He
Route IV PO IV PO IV PO IV PO
N' 4 3 3 3 3 3 3 3
Dose(mg/kg) 2.5 10 2.5 4 5 10 5 10
ClAtnUmin/kg) 7.7 1.0 - 22 13 -- 3 -- - -- 13 2
Vss'(Ukg) 4.9 1.9 - 0.33 0.25 - ,1.4 0.2 - 1.4
0.2
Al/C1(min*mg/mL) 279 53 37 20 139 '77 0.4 296 46 65
20 381 65 160 13
C e(nemL) 3816 509 212 3.2 1.6 3794 1580 4/ 98 438 814 255
3349 686 1262 362
Fr(%) 3.3 0.2 11 21
`t,
a Numbers of rats. Systemic clearance. C Volume of distribution following
intravenous dosing. d
Area under the curve following intravenous dosing, integrated drug
concentration with respect to
time and integrated drug concentration with respect to time following oral
dosing. Maximum
plasma concentration following intravenous dosing. I Percent oral
bioavailability.
[00428] Modifying Substituted Methoxybenzoyl Aryl Thiazole (SMART)
Molecules to
Improve Oral BiOavailability. Many established tubulin targeting anticancer
drugs like taxanes
.. and vinblastine require intravenous administration because of low oral
bioavailability. Oral
bioavailability is a complex parameter involving many chemical and
physiological processes,
such as solubility, permeability, and metabolic stability. The solubility of
these tubulin inhibitors
was improved by inserting an amino linker between the "A" and "B" rings as in
5a-d (Figure 6),
Table 3 demonstrates that the NH bridged compounds (5a-c) had similar potency
(35-: 65 nM) as
lh with increased solubility (15 and 19 vg/mL for 5a and Sc, respectively
(Figure 18), but they
are over 20 fold more active than ABT-751 (Table 3 and Figure 19 for the
structure of ABT-
751).
[00429] Rat pharmacokinetic studies were performed to study whether
these new
compounds exhibited improved bioavailability compared to compound 1h (Table
4). The data
190
CA 3030689 2019-01-18
clearly showed that 5Hc (HCI salt of Sc) exhibited more than 4.3-fold
increased exposure (AUC)
by the oral route as compared to lh, suggesting that improved aqueous
solubility by the amino
linker successfully improved oral bioavailability. In addition, the maximal
concentration (Cmax)
of 5Ha and 5Hc by oral administration was 814 and 1262 ng/mL, respectively.
While Cmax of
lh was only 212 ng/mL. Overall, the bioavailability of 5Ha and 5Hc was
increased from 3.3% of
lh to 11% and 21%, respectively (Table 4). Compound 5Hc exhibited moderate
clearance,
moderate volume of distribution, and acceptable oral bioavailability. This
data suggested that
these new synthesized amino linked compounds have the potency and PK profile
to be developed
as a new class of orally bioavailable antitubulin agents.
Example HD: in vitro Tubulin Polymerization inhibition by compounds of the
invention.
[00430] In Vitro Tubulin Polymerization Assay. Bovine brain tubulin
(0.4 mg, >97%
pure) (Cytoskeleton, Denver, CO) was mixed with 10 M of the test compounds and
incubated in
100 pi of general tubulin buffer (80 mM PIPES, 2.0 mM MgCl2, 0.5 mM EGTA, and
1 mM
GTP) at pH 6.9. The absorbance of wavelength at 340.nm was monitored every 1
min for 20 min
by the SYNERGY 4 Microplate Reader (Bio-Tek Instruments, Winooski, VT). The
spectrophotometer was set at 37 C for tubulin polymerization.
Results:
[00431] The inhibition of tubulin polymerization by selected potent
compounds le, 2j,
66a, and 5a was investigated by all three design strategies (alternative B-
rings, novel linkers, and
solubilizing moieties) and compared with lh. Bovine brain tubulin (> 97% pure)
was incubated
with the individual compounds (10 1.1.M) to test their effect on tubulin
polymerization (Figure
20a). After 20 min incubation, tubulin polymerization was inhibited 47% by lh,
as compared to
.. vehicle. Both le and 2j inhibited 64% of polymerization at 20 min with
different inhibition
patterns. Compounds 5a and 66a provided greater inhibitions as 78% and 81%,
respectively.
These data suggest that these compounds exhibit strong antitubulin
polymerization activity that
corresponds well with their cytotoxicity.
[00432] The inhibition of tubulin 'polymerization by compound 5c by
binding with
colchicines binding site and compared with compound lh is demonstrated in
Figures 20b and
20c.
191
CA 3030689 2019-01-18
Example 11E: Novel Anti-tubulin Compounds Overcome P-Glvcoprotein Mediated
Multidrug Resistance.
[00433] The P-glycoprotein (P-gp) system appears to be a primary
physiological
mechanism of multidrug resistance (MDR) which acts as an ATP-dependent drug
efflux pump,
actively removing a variety of structurally diverse cytotoxic compounds.
Enhanced efflux of =
these compounds reduces their intracellular accumulation and so reduces their
cytotoxicity.
Therefore, novel compounds which are not susceptible to drug resistance could
be of high
therapeutic and economic value, hi addition to P-gp, clinically used
antitubulin agents have other
resistance mechanisms such as changes in microtubule dynamics and mutations in
P-tubulin
which are known to limit sensitivity to the taxanes. The anti-tubulin
compounds of the invention
were tested against an ovarian cancer cell line OVCAR-8 (parent) and P-gp over-
expressing
NC1/ADR-RES cell line (Tables 5A, 5B).
Results:
Table 5A. Antiproliferative Activity of Selected Compounds against P-gp over-
expressed MDR
cell lines.
1050 (nM) Resistance
Compound
OVCAR-8 NCl/ADR-RES factor
lc 33 3 13 0.8 0.4
2j 34 2 14 1 0.4
66a 10 3 4 2 0.4
2r 26 2 11 2 0.4
5a 46 6 27 0.6
5b 28 21 0.8
Sc 44 3 25 6 0.6
lh 35 2 13 1 0.4
paclitaxel* 4.7 0.1 6263 634 1333
vinblastine 3.9 0.1 582 57 149
colchicine 17 1 I113 79 65
192
CA 3030689 2019-01-18
[00434] Notably, the anti-tubulin compounds of the invention demonstrated
equipotent
antiproliferative effects against OVCAR-8 and NCl/ADR-RES cell lines,
suggesting that they are
not P-gp substrates and that they function in a P-gp-independent manner. This
feature is distinct
from that of paclitaxel, vinblastine, and colchicine in NC1/ADR-RES cells.
-
Table 5B. Antiproliferative activity of selected phenyl-amino thiazole
compounds
IC50 SEM (nM)
B16-F1 A375 DU 145 PC-.3 LNCaP PPC-1
5a H 65 12 45 8 70 4 57 3 51 1 54 1
5Hb 4-CH3 ND* ND 35+1 38 2 35 1 36 1
5c 4-F ND ND 63 1 43 1 41 1 37 1
-5(1 4-C1 ND 25 7 73 1 33 1 45 1 36 1
lh 55 5 28 5 71 4 21 1 28 4 43 5
ABT-751 - 2127 351 1111 108 839 719 786 89 658 117 701 307
The phenyl amino thiazole compounds 5a, 5Hb, Sc and 5d demonstrated potent
activity in a
number of prostate cancer cell lines. Unexpectedly, the phenyl amino imidazole
compound 5e
demonstrated no activity (ICso > 1000 nM in LNCaP, PC-3,. DU-145, and PPC-1)
in these
' prostate
cancer cell lines. The positive controls for this experiment were 55 and 17ya
which
demonstrated IC50 values between 7.5 nM and 24.1 nM in the same cell lines
(Table 5C).
= 15
[00435] Table 5C
Table 5C. ICso SEM (nM)
B16-F1 A375 DU 145 PC-3 LNCaP PPC-1
5a 65 12 45 8 70 4 57 3 51 1 54 1
5Hb ND ND 35 1 38 2 35 1 36 1
Sc ND ND 63 1 43 1 41 1 37 1
5d ND 25 7 73 1 33 1 45 1 36 1
lh 55 5 28 5 71 4 21 1 28 4 43 5
ABT-751 2127 351 1111 108 839 719 786 89 658 117 701+307
d ND ND >1000 >1000 >1000 >1000
ND ND >1000 >1000 >1000 >1000
193
CA 3030689 2019-01-18
Se ND ND >1000 >1000 >1000 >1000
55 ND ND 24 6 12 1 13 1 15 1
17ya ND ND 11 1 5 2 8 2 8 1
[00436] A new series of tubulin polymerization inhibitors with
acceptable oral -
bioavailability and equi-potent activity in multidrug resistant tumor cell
lines has been
discovered. Medicinal chemistry efforts starting from optimizing SMART
compound lh.
Chemical modifications of different substituted aryl in "B" ring and linkages
between "B" and
"C" rings were investigated based on biological evaluation against cancer
cells in vitro. SAR
studies revealed that optimal "B" rings include .pyridine (lc), thiophene
(1f), and furan (1d)
which maintain excellent in vitro potency. Replacing carbonyl linker with
cyanoimine (2j)
between "B" and "C" ring will increase the activity. Structure modifications
to increase aqueous
solubility and bioavailability were performed. Introducing an amino between
"A" and "B" rings
gave us compounds 5a-c, which showed similar in vitro antiproliferative
potency against tested
cancer cells as well as MDR(+) and MDR(-) cell lines, furthermore, the
solubility and in vivo
bioavailability were improved greatly over those of the lh. Therefore, these
new anti-tubulin
compounds represent a new family of compounds that may be very useful in the
treatment of
cancer.
194
CA 3030689 2019-01-18
o P-74392-PC I
,
.
co
0
w
0
0,
co EXAMPLE 12
l0
1J ANTIPROLIFERATIVE ACTIVITY OF COMPOUNDS OF
THIS INVENTION.
0
I-`
l0
I
0
I-` 5 [004371 . The antiproliferative activity of analogs
prepared by the methods of the invention are shown in Tables 6 and 6A.
1
I-`
CO
Table 6
leso SEM (nM)
ID LNCaP PC-3 DU 145 PPC-1 A375 816-F1 WM164
MES- MES- OVCAR NCl/AD
SA SA/Dx5 -8 R-RES
Paclitaxel 1.7 4.8 5.1 2.3 12 17
2.7 6.6 4.7 6263
Vinblastine 1.1 2.1 1.8 1.1 1 4.7
1.4 16 3.9 582
Colchicine 16 11 ' 10 ' 20 20 29
= 8.4 22 17 1113
1k 101 - 101 140 ' 84 100 ' 245
220
2k 6 r 13 12 8 33 43
11 19 34 12
2m 19 - 8.7 6.9 6.2 11 21
2n 101 131 143 99 210 . 290
2o 65 - 73 121 ' 73 38 42
2p >10000 ' 2385 1899 1079 2200
16560 =
2q >10000 >10000 >10000 >10000 >20000 >20000 .
5c-HCI 53 53 70 43
6d 703 908 1637 929
*ND: not determined .
Table 6A
195
P
,...,
IC50 (nM)
0
0,
cc) Structure ID LNCaP PC-3 DU 145 P PC-1 A375
B16-F1 WM164 MES- MES- OVCAR NCICAD
to
IQ
SA SA/Dx5 -8 R-RES
0
1-`
l0 0 i 8 346 704 580 230 318 570 404
o1 /
1-`
1-`
CO S ./1=1 H
HN
_
9 -10000 -10000 -10000 -10000 ¨ --
H Cµo
*
. liSi CH-CH3
I
¨ _______________________________
' 658 786 839 701 :1111 2127
661 .
-N !
NH 1
/
OzS
I
I
,..,.0
1
196
o
0
0
Structure ID LNCaP PC-3 DU 145 PPC-1 A375
B16-F1 WM164 MES- MES- OVCAR NCl/AD
CO
SA SA/Dx5 -8
R-RES
0
0 11 >10000 >10000 -10000 -10000 3470 4900
4700
0
1-`
CO
0 12 >10000 I >10000 >10000 I >10000 >10000
>10000
13 >10000 >10000 >10000 >10000 >10000 >10000 >10000
0 *
0 14 >10000 >10000 >10000 >10000 >10000 -
>10000
197
ctt:
Structure ID LNCaP PC-3 DU 145 PPC-1 A375
B16-F1 WM164 MES- MES- OVCAR NCl/AD
to
SA SA/Dx5 -8 R-RES
o 16 >10000 >10000 >10000 >10000 15200 6900
0
F
p_krIk
F
17 2100 1900 2600 1300 4300 9800
r
>
F
/1
H3C. 18 ¨10000 ¨10000 ¨10000 ¨10000
;0 0
O-Ch
0
\-27
X 3
=
19 >20000 >20000 >20000 >20000 >20000 >20000
\ 7
198
o
0
0
o
co Structure ID LNCaP PC-3 DU 145 PPC-1 A375
B16-F1 WM164 MES- MES- OVCAR NCl/AD
SA SAIDx5 -8
R-RES
0
20 1452 >10000 642 633 2300 3100
1300
0
1-`
o
CO
21 314 403 435 216 383 924 408
N NH
HN
22 >20000 >20000 >20000 >20000>20000 >20000
23 ¨10000 ¨10000 ¨10000 ¨10000
_______________________________________________________________________________
6 N 0 CF0
1
199
o
0
0
Structure ID LNCaP PC-3 DU 145 PPC-1 A375 B16-F1 I WM164 MES- MES-
OVCAR NCl/AD
co
to
SA SA/Dx5 -8
R-RES
o
1-`
24 >10000 >10000 >10000 >10000 >10000 >10000 >10000
o
1-`
1-`
\ 0
CO
\o 25 48 44 24 13 20 38
0
0
N
26 23 16 16 15 11 14
0
N 0¨
H2N
200
o
0
Structure ID LNCaP PC-3 DU 145 PPC-1 A375
1316-F1 WM164 MES- MES- OVCAR NCl/AD
0
co
SA SA/Dx5 -8 R-RES
29 1788 >10000 >10000 >10000 >10000 >10000
0
1-`
CO
30 >10000 >10000 >10000 >10000 >10000 >10000
0
NC OMe 32 1664 2291 4601 1170 2700
>10000 , 2600
OH
S N OMe
110
201
o
Structure ID LNCaP PC-3 DU 145 PPC-1 A375
B16-F1 I WM164 MES- MES- OVCAR NCl/AD
CO
SA SA/Dx5 -8
R-RES
0
H 33 >2000 >2000 >2000 >2000 9800 >20000
0
1-`
CO OH
H3c`o 34 >10000 >10000 >10000 >10000 >10000 >10000
>10000
O-CH
Hp¨o 35 1500 40100 21900 15000
H3C
0
H3d'
0 0-0-1,
\CH,
H30-0 39 4300 32500 16800 21400
=
HC
0 -OH
202
o
0
Structure ID LNCaP PC-3 DU 145 PPC-1 'A375 B16-
F1 WM164 MES- MES- OVCAR NCl/AD
CO
SA SA/Dx5 -8
R-RES
0 40 13400 19600 18400 6200
0
1-`
CO
41 15750 18170 17040 >20000
110
41*
0
40,
42 ___________________________________ 43590 23790 24880 >20000
B
0
43 12690 14720 17210 >20000
203
Structure ID LNCaP PC-3 DU 146 PPC-1 A375
B16-F1 WM164 MES- MES- OVCAR NCl/AD
co
to
SA SA/Dx5 -8 R-RES
ocH3 17Y 12 10 17 21 17.35 32.94 12.08
0
o ocH, a
HN N OC H3
CO
HN 441
Ph 17y 233.7 148.3 592,1 208.9 481.2 538.7 1
467.6
o=s-c
oc 22
N /NJ? 110
/ I OC
OCH,
16x 1068 2628 5917 4575 1800
1390 1700
aa
s
:2
16x >10000 >10000 >10000 >10000 >10000 >10000 >10000
HF,0
aa
"
es.ii¨Nsp,,H 0-CH,
,
204
EXAMPLE 13
BIOLOGICAL EVALUATION OF ISOOUINOLINE DERIVATIVES OF THIS
INVENTION
Cell culture.
= 5 [00438] LNCaP, PC-3, DU-145, PPC-1, MES-SA, and MES-SA/DX5
were originally
obtained from ATCC (Rockville, MD). All cells obtained from ATCC were
immediately
expanded and frozen down such that all cell lines could be restarted every 2-3
months from a
frozen vial of the same batch of cells. For the in vivo xenograft studies, PC-
3 was authenticated at
Research Animal Diagnostic Laboratory (Columbia, MO) within four months before
studies.
Inter-species contamination was tested by PCR and the identity of the cell
lines was verified by
generating -a genetic profile. MES-SA and MES-SA/DX5 were maintained in
McCoy's 5A
Medium containing 2 mM L-glutamine supplemented with 10% fetal bovine serum
(FBS). All
other cells were maintained in RPMI-1640 medium with 2 mM L-glutamine and 10%
FBS.
Growth Inhibition Assay.
[00439] The cytotoxic or antiproliferative activity of test compounds
was investigated in
' several cell lines using the sulforhodamine B (SRB) assay. Cultured cells
were plated into 96-
well plates and incubated with medium containing different concentrations of
the test compounds
for 96 h. Cells were stained with SRB solution. The optical density was
determined at 540 nm on
a microplate reader (Dynex Technologies, Chantilly, VA). Plots of percent
inhibition of cell
growth versus drug concentration were constructed, and the concentration that
inhibited cell
growth by 50% relative to the untreated control (IC50) was determined by
nonlinear least squares
regression using WinNonlin software (Pharsight Corporation, Cary, NC).
Cell Cycle Analysis.
[004401 Cell cycle distribution was determined by propidium iodide
(PI) staining. Treated
cells were washed with PBS and fixed with 70% ice-cold ethanol overnight.
Fixed cells were
then stained with 20 mg/mL of PI in the presence of RNase A (300 g/mL) at 37
C for 30 min.
Cell cycle distribution was analyzed by fluorescence-activated cell sorting
(FACS) analysis core
services at the University of Tennessee Health Science Center, TN.
205
CA 3030689 2019-01-18
In Vitro Metabolism Studies.
[00441] For both phase I, the incubation mixture, in 65 mM potassium
phosphate buffer
(pH 7.4), consisted of 1 mg/mL liver microsomal proteins, 3 mM NADPH, and 0.5
M test
compound. The concentration of methanol (used for dissolving the substrate)
was 1 % (v/v).
Total volume of the incubation was 200 ptI. and the reaction mixtures were
incubated at 37 C.
To generate the stability curves for test compounds different incubations were
stopped at ID, 20,
30, 60, and 90 minutes for analysis of compounds remaining. All reactions were
stopped by the
addition of 200 1.. ice-cold acetonitrile. Subsequently, the samples were
then centrifuged at 3000
g for 5 min and supernatant was analyzed by LC-MS/MS.
Pharmacokinetic Studies in Mice.
[00442] Male ICR mice (5-6 weeks, 20-25 g) were used. For 6a, 6b, and
6c a dose of
5mg/kg was administered via the i.v., i.p., and p.o. route. Iv. doses were
administered via the tail
vein. Oral doses were administered by gavage. At each time point, three to
four mice were
.. euthanized by isoflurane (Baxter Healthcare, Deerfield, IL) and blood
samples (up to 600 lit
each) were taken from the posterior vena cava. Plasma samples were stored at -
20 C prior to
analysis. Plasma proteins were precipitated by the addition of acetonitrile
(150 L, containing the
internal standard) to 100 1., of mouse plasma. Samples were vortexed and then
centrifuged at
8000g for 10 min. The supernatant was transferred to a clean vial for
injection into the mass
spectrometer for analysis.
In Vivo Antitumor Efficacy Study.
[00443] PC-3 cells (2.5x106 cells/site) plus Mairigel (BD biosciences,
San Jose, CA) were
injected subcutaneously into flanks of male nu/nu mice. Tumor size was
measured using calipers
every 2-4 days and calculated as V it / 6 x (length) x (width)2. When tumors
reached a volume
of approximately 100-150 nun3, drug treatment was initiated. The control group
was treated with
vehicle (20% Captex200 in Tween80). During the treatment, tumor size and body
weights were
measured every 2-4 days.
White Blood Cell Counting.
[00444] Whole blood was obtained from nude mice at the end of efficacy
study. To count
white blood cells (WBC) using a hemacytometer, 10 j.tL of whole blood sample
was diluted with
206
CA 3030689 2019-01-18
the 190 1.tL of 2% acetic acid. With proper light adjustment, the leukocytes
appeared as dark dots
on the hemacytometer. WBC in each sample was counted twice within one hours
following
dilution and average was calculated.
Results
Table 7. Anticancer efficacy of isoquinoline compounds in different cancer
cell lines and MDR cell lines
mediated by P-glycoprotein
tc5c, (nM)
6a 6b 6c Vinblastine
Docetaxel
LNCaP 80.6 17.1 98.1 17.9 38.3 9.7 ,
3.4 0.9 4.7 1.3
PC-3 = 64.4 12.2 71.8 9.1 25.6 8.3 1.4
0.3 6.3 0.4
DU-145 91.7 10,2 113.4 21.4 46.6 13.8
2.6 1.0 5.2 1.0
PPC- I 60.6 3.4 47.9 10.0 27.7 4.5 1.1
0.4 2.7 1.0
P-gp
MES-SA 78.2 1.8 129.8 38.0 35.6 2.8 2.3
0.8 5.9 1.1
MES-SA/DX5 119.4 0.4 177.8 32.8 59.2 0.1
45.7 5.3 76.4 8.7
Resistance factor 1.5 1.4 1.7 20 13
NOTE: P-gp is over-expressed in MES-SA/DX5. The resistance factor (RF) was
calculated as the ratio
of 1050 values for the resistant cell subline to that of the parental cell
line. All experiments were
performed at least in three replicates.
=
Table 8. Compound 6a, 6b, and 6c arrested PC-3 cells in G2M phase.
G2M phase arrest EC50(nM)
6a 53.4
6b 91.9
6c 23.3
15
=
207
CA 3030 6 8 9 2 0 1 9 -0 1-1 8
Table 9. Summary of half lives (phase I pathway) of 6a, 6b, and 6c in mouse,
rat, hamster,
rabbit, guinea pig, dog, monkey, and human liver microsomes.
T V2 (min)
6a 6b 6c
_
Mouse 3.4 10 , 13
Rat 12 9 14
Hamster 6 11 20
Rabbit 17 16 16
Guinea pig 15 15 , 8
Dog - 13 30 29
Monkey 16 13 9
Human 32 40 47
Table 10. Summary of pharmacokinetic properties of compound 6a, 6b, and 6c in
mice.
6a 6b 6c
'-
1
143C0 -.' OCH3
"3 H
MW 410.5 359.4 338.4
IV CL (mL*Min'Ike) _ 5mg/kg 51 14 30
IV Vd (1,*kg-1) 5mg/kg 2.3 1.1 1.8
W C.õ (ng/mL) 5mg/kg 678.4 1500 1100
IP AUC (mini.tg/mL) 5mg/kg 59 218 55
IF Bioavailability Fi% 60 60 33
PO C.. (ng/mL) 5mg/kg 6.7 50 50
AUC (min*pg/mL) _ 5mg/kg 5 7 4 _
PO Bioavailability _ Fp.% 5 2.1 2.7
_
_
[00445] Efficacy and tolerability of 6b and 6c was measured in xenograft
models after i.p.
injection (Figure 34). PC-3 xenografts were treated with vehicle (qd), 6b (40
mg/kg, qd), or 6c
(40 mg/kg, qd) for 3 weeks. Dosing vehicles were composed of 20% Captex200 in
Tween80. The
tumor volumes (mm3) were plotted against time and are the means SD from
eight animals. The
tumor volumes and survival rates or body weights are shown in Figure 34A. The
liver size (g) of
each nude mouse was measured after 3 weeks treatment and is shown in Figure
34B. The
208
'
CA 3030689 2019-01-18
number of white blood cells was counted in whole blood collected from animal
after 3 weeks
treatment and is shown in Figure 34C.
=
EXAMPLE 14
ANTIPROLIFERATIVE ACT1VTTY OF SELECTED ABI COMPOUNDS OF THIS
INVENTION
Cell Culture Cytotoxicity Assay
Materials and Methods
[00446] The antiproliferative activity of the ABI compounds in three
melanoma cell lines
(A375 and WM-164, human melanoma cell line; B16-F1, mouse melanoma cell line)
and four
human prostate cancer cell lines (LNCaP, DU 145, PC-3, and PPC-1) were
studied. All these cell
lines were purchased from ATCC (American Type Culture Collection, Manassas,
VA) except the
PPC-1 cell line. MDA-MB-435 and MDA-MB-435/LCCMDR1 cells were kindly provided
by
Dr. Robert Clarke at Georgetown University School of Medicine, Washington, DC.
Melanoma
cells were cultured in DMEM (Cellgro Mediatech, Inc., Herndon, VA) and
prostate cancer cells
were cultured in RPMI 1640 (Cellgro Mediatech, Inc., Herndon, VA) supplemented
with 10%
FBS (Cellgro Mediatech). Cultures were maintained at 37 C in a humidified
atmosphere
containing 5% CO2. 1000 to 5000 cells were plated into each well of 96-well
plates depending on
growth rate and exposed to different concentrations of a test compound for 48
h (fast growing
melanoma cells) or 96 h (slow growing prostate cancer cells) in three to five
replicates. Cell
numbers at the end of the drug treatment were measured by the sulforhodamine B
(SRB) assay.
Briefly, the cells were fixed with 10% trichloroacetic acid and stained with
0.4% SRB, and the
absorbances at 540 nm were measured using a plate reader (DYNEX Technologies,
Chantilly,
VA). Percentages of cell survival versus drug concentrations were plotted, and
the IC50
(concentration that inhibited cell growth by 50% of untreated control) values
were obtained by
nonlinear regression analysis using GraphPad Prism (GraphPad Software, San
Diego, CA).
Results
[00447] The results of the in vitro antiproliferative activities of the
compounds of this
invention using three melanoma cell lines (one murine melanoma cell line, B16-
F1, and two
209
CA 3030689 2019-01-18
human metastatic melanoma cell lines, A375 and WM-164) and four human prostate
cancer cell
lines (LNCaP, PC-3, Du 145, and PPC-1) are summarized in Tables 11-13.
=
210
CA 3030689 2019-01-18
P-74392-PC I
P
,....,
0 [004481 Table 11. In vitro growth inhibitory effects of
compounds without A ring substitutions.
ui
0 .
0,
co
to
m
0
1-` 1C30
(nM) .
to
o1 ID Structure R A375 B16-F1 WM164
1-`
LIVCaP PC-3 Du 145 .PPC-1
1
,
1-` ¨
co 4111 12aa 3,4,5-(0Me)3 160 120 10
152 288 196 133
12ab 4-0Me >10000 >10000 > 1
0000 >10000 >10000 >10000 >10000
_
N NH
12ac 3-0Me >10000 >10000
>10000 >10000 >10000 >10000 >10000
'
,
\-_=/ 12a 3,5-(0me), 2800 5400 2100
3611 3274 2590 2129
0= 12ae 3,4-(0Me)2 >10000 >10000
>10000 >10000 >10000 >10000 >10000
r) 12a1 4-F 580 930 630 613
2197 846 575
R 12ag 3-F >10000 >10000
>10000 >10000 >10000 >10000 >10000
12ah 4-Me >10000 >10000
>10000 >10000 >10000 ' >10000 >10000
. . ,
12a 3-Mc >10000 >10000 >10000 >10000
>10000 >10000 >10000
.
.
12aba 4-0Me >10000 >10000
>10000 >10000 >10000 >10000 >10000
o.,
,
.
1 I 12aaa 3,4,5-(0Me)3 >10000
>10000 >10000 >10000 >10000 >10000 >10000
R
N
1 2 H >10000 >10000
>10000 >10000 >10000 >10000 >10000
R 0.6-C
1 Ox 4-NO2 >10000 >10000
>10000 ' >10000 >10000 >10000 >10000
-4
10j 4-0B n >10000 >10000 >10000
>10000 = >10000 >10000 >10000
_ -
=
211
[00449] From Table 11, compounds 12aa-12a1 showed moderate activity
with IC50 values
in the M range (average of all seven cell lines). The most potent compound of
this series was
12aa with an average IC50 value of 160 nM. The removal of one of the methoxy
groups from the
3,4,5-trimethoxy on the C ring (12ad, 12ae) led to a significant loss of
activity (IC50 >10 M for
12ae and an average IC50 of 3.1 tt.M for 12ad). Compound with 4-fluoro on the
C ring (12af) also
showed relatively good activity (IC50 = 0.91 M), a finding that has an
important implication,
because replacing the trimethoxy moiety with a 4-fluoro group may provide good
activity and
improved metabolic stability. The position of the fluorine on the C ring
was_critical for activity
because a shift from 4-fluoro to 3-fluoro resulted in a total loss of activity
(IC50 >10 M for 12ag
compared with 0.91 M for 12a1). This result suggeisted that a potential
hydrogen bond donor is
present close to the 4-position of this ring.
[00450] As clearly indicated in Table 11, the positions of the A and C rings
were critical. A simple
shift of the C-ring moiety from position 4 to position 1 in the imidazole ring
(B ring) resulted in total
loss of activity (IC50 >10 M for 12aba, 12aaa, 10a, 10x, 10j).
212
CA 3030689 2019-01-18
o
P-74392-PC1 .
,
u.,
0
u.,
0
0,
co
to [00451] Table 12. /n vitro growth inhibitory effects of
compounds with substitutions on A ring.
m
o .
I-,
l0
-
oI
I-, IC50
SEM (nM)
1 .
-
CO ID RI R2 = -
A375 B16-F1 WM164 LNCaP PC-3
Du 145 PPC-1 OVCAR-8 NO1/ADR-
RES
_
fr.!? 12ba 4-F 3,4,5-(0Me)3 205 19 ' 320 41 73t8
98t2 169 12 -132 24 81 1
_
12ca 4-0Me 3,4,5-(0Me)3 30t5 I 08t1 2 31 4
31t1 45 1 4810.5 34 -0.3
12c6 443N4e 4-F 31 5 63 7 - 28 3 l8 2
31t2 11 38 29t1
\=1
12da 4-Me 34,5-(0Me)3 9 2 46t5 - 8t2 12 1
9t0.4 15 0.5 I11t0.1
-
12db 4-Me 4-F 143 13 222 10 156 19 15+2 56t3
78t5 54t1
0 = x" 12db-11C1 108t I 1 297 23 1121-9
VD \11) ND ND
F12
-
3,5-(0M02- '
12dc 4-Me 105 187 123 134 127
174 110
4-0H
3,4,5-
12ea 3,4,5-(0Me)3 4800 >10000 >10000 >10000 >10000 >10000
>10000
(0Me)3
3.4,5-
12eb 4-F >10000 >10000 >14000 >10000 >10000 >10000 >10000
(0Me)3
12fa 4-CI 3,4.5-(0Me)3 43t5 168 14 26 3
2411 15i I 360.4 261-0.2 47 19
12113 4-CI 4-F , 52 4 73 6 741-9 49t2 'It2 55 1
52i1
-
L
131a 4-CI 3,4,5-(OH)3 3900 1810 2100
10000 10000 10000 >10000
12ga 4-N(Me)2 3,4,5-(0Me)3 82 9 361 29 80 11
58t2 92 4 95 1 67 0.7
12gb 4-N(Me)2 4-F 56t7 129 11 6203 57 6
8 I -3 72 0.4 45 0.3
3,4-
12ha 3,4,5-(0Me)3 113 14 1400 200 191t18 121 10 /03 7 168 15
117 1 -
(0Me)2
3,4-
1211b 4-F 10000 4210 1400 2533
10000 10000 2172 48
(01%402
1
213
ri
P-74392-PC1
u.) .
0
u.) 121a 2.-CF3 3,4 .5-(0Me)3 >10000
>10000 >10000 >10000 >10000 >10000 >10000
0
co 121b 2-CF3 4-F - >10000 >10000
>10000 ' >10000 >10000 >10000 >10000
(.0
3,4,5-
IQ 13ea 3,4,54011)2 >10000 >10000 >10000 >10000
>10000 >10000 >10000
o (OH)
1-`
t.0 1 12ja ' 4-013n 3,4,5-(0Me)3 5200 10000 -
5500 2786 10000 10000 2844
o ,
1-` 12jb 4-0Bn 4-F 93t8 117 16 90 12
44 7 79 0.4 60 3 43 0.2
i
1-`
co 121ca 4-01{ 3,4,5-(0Me)3 1600 2400 -1
1800 ND ND ' ND - ND
12kb 4-0H 4-F 10000 >10000 >10000 10000 >10000
>10000 >10000 -
¨ ___________________________________________________
3-0H, 45-
12kc 4-0H 10000 5600 6400
(0Me)2
121a 4-Br 3,4,5-(0Me)3 32 74 --I 36
34 _____ 36 = 49 33
_
_______________________________________________________________________________
__________________________
12pa 4-CF3 3,4,5-(0Me)2 163.1 468.7 175
134 127 174 110
_
_______________________________________________________________________________
__________________________
13 ha 3,4-(OH)2 3,4,5-(011)3 ' >10000 ' >10000 >10000
ND ND ND ND
. . .
.
Colchicine 2033 ' 29t5 ND 1634 ' 11 1
' 10 .2 ' 20t1 '
ND- not determined
= -
,
'
- .
214
[00452] From Table 12 compounds with 3,4,5-trimethoxy and 4-fluoro
substitutions on
the C ring showed good activity with different substitutions on the A ring.
These compounds
demonstrated excellent antiproliferative activity with IC50 values as low as
8.0 nM on WM164
cell line (12da). In general, compounds incorporating a single substituent on
the para-position of
the A ring were more potent as can be seen from the activities of 12ca, 126,
12da, 12db, 12fa,
12Th, 12ga, and 12gb (IC50 = 7.9-110 nM). 12db-HC1 salt (IC50 = 172 nM) showed
slightly
diminished activity compared with the corresponding free base 12db (IC50 = 109
nM).
Compound 12113 (IC50 = 63.7 nM), with a single halogen substituent in the para-
position of the A
and C rings, demonstrated potent and was devoid of a methoxy moiety. Compounds
with 3,4,5-
trimethoxy substituents on the A ring lost activity completely (IC50 > 10 M
for 12ea, 12eb),
suggesting very different binding environments near the A ring and C ring.
Removal of the 5-
methoxy. substituent from the A-ring improved activity significantly (IC50 =
330 nM and >10 AlVI
for 12ha, 12ea respectively). Demethylation of the 3,4,5-trimethoxy decreased
activity sharply
from 43 nM (12fa) to 3.89 ialvl (13fa). Similar results were observed for 136,
12ka, 12kb, and
13ha due to the demethylation of subsituents on either the A or C ring.
Electron-donating groups
(4-methoxy, 4-dimethylamino, 4-methyl) and electron-withdrawing groups (4-
chloro, 2-
trifluoromethyl) on the A ring did not show substantial differences in
activity. The introduction of
a trifluoromethyl group at the ortho position of the A ring caused complete
loss of activity (IC50
>10 uM for 12ia, 12ib). The presence of a benzyloxy group at the para position
of A ring (IC50 =
75 nM for 12jb) resulted in a 440-fold increase in activity when compared with
the para-hydroxy
compound 12kb (1050,-33 ItM). It is worthwhile to note that compound 12jb,
with the 4-fluoro in
the C ring, has better activity than does its counterpart 12ja, which has a
3,4,5-trimethoxy group
in the C ring (1050 is 75 nM for 12jb, and 7.3 RM for 12ja).
215
CA 3030689 2019-01-18
ri
P-74392-PC I
u.)
0
(.,.)
0
co
io [004531
Table 13. In vitro growth inhibitory effects of compounds with protection
on B ring.
m
o .
1-`
tO
oI
IC50 SEM (nM)
1-` Structure ID RI itz R3 -
1
1-` A375
B16-F1 WM164 LNCaP PC-3 Du 145 PPC-1
co
4 llab H 4-0Me SO2Ph 44
>10000 >10000 4 >10000 >10000 >10000 >10000 ' >10000
Ilac H 3-0Me 4 502Ph
>10000 >10000 >10000 - >10000 >10000 >10000 >10000 4
llah H 4-Me S02Ph
>10000 >10000 >10000 - >10000 >10000 >10000 >10000 .
,._.
I 1 af H 4-F S02Ph . 630 72
946 86 5961-61 573 ' 2233 846 575
-=
hag H 3-F SO2Ph
>10000 >10000 >10000 >10000 >10000 >10000 >10000
¨
I Icb 4-0Me 4 4-F 502Ph 36 5
71t8 43 6 31 2 33 2 520 32 0.7 '
R1 I Idb 4-Me 4 4-F 502Ph ' 113 14 287 31 107
14 55 3 80 1 80 1 57 1
I Ilea 3,4,5-(0Me); 3,4,540Me)3 502Ph
>10000 >10000 > I 0000 >10000 >10000 >10000 >10000
-,i-
Ileb 3,4,5-(0Me)3 4-F 502Ph
3840 >10000 ' >10000 >10000 >10000 ' >10000 >10000
RI. N "1-:-.N
lltb 4-CI 4-F SO2Ph
88+9 I07 12 70t6 48 1 76 2 64 I i 54 1
0= 11 ga 4-N(Me)2 3,4.5-(0Me)3 S02Ph 162 13
1200 90 308+32 62 2 93 5 99 2 72 0.4
--- I lgb 4-N(Mc)2 4-F SO,Ph 55 7 242t26 56 4 561-6
83 3 74 0.5 48 0.3
_
llha 3,440Me12 3,4,5-
(0Me)3 SO2Ph 192 15 9701-68 139-1-15 1 I 4 6 197 9 144 29
11712
N2
I Ihb 3,4-(0Me)2 4-F SO2P11
960 59 2000 400 1400 30 1915 77 10000 4 3312 14411-49
Ilia 2-CF3 3,4,5-(0Me)3 5 02Ph >10000
>10000 >10000 >10000 >10000 ' >10000 >10000
1 1 ib 2-CF3 4-F S 02Ph >10000
>10000 >10000 >10000 >10000 >10000 >10000
I ljb 4-013n 4-F S02Ph 64 7 110 15
48 5 4 35 I 75 0.5 58 1 38 0.2
12dab 4-Me 3,4,5-(0Me)3 Me 32 134 40
32 46 36 28
12cba 4-0Me 4-F 4 Mc >10000
>10000 >10000 >10000 >10000 >10000 >10000
12daa 4-Me 3,4,5-(0Me)2 C1-121,11
683.2 465.8 1501 777.9
12gba 4.N(Me)2 4-F S02PhOMe -100 -100 -100
73.2 44.14 129.4 63.4 .1
_.
'
216
[00454] From Table 13, compounds with a phenylsulfonyl protection group
attached to the
nitrogen of the imidazole ring (llcb, lldb, 11M, llga, llgb, llha, lljb) were
also very active
with IC50 in the nM range (Table 13). Generally the activities of these
compounds are
comparable to their corresponding unprotected counterparts as exemplified by
comparing the
activities of llcb (43 nM), Ildb (111 nM), 11th (72 nM), llga (285 nM), 11gb
(87 nM), llha
(268 nM), and lljb (61 nM) with their corresponding unprotected counterparts
12cb (36 nM),
12db (109 nM), 12113 (64 nM), 12ga (131 nM), 12gb (72 nM), 12ha (330 nM), and
12jb (75
nM). Other compounds (11ab-llag, Ilea, lleb, llhb, Ilia, and llib, 1-50 p.M)
were generally
much less active, also in line with their counterparts (12ab-12ag, 12ea, 12eb,
12hb, 12ia, and
12ib, 1-50 M).
EXAMPLE 15
ACTIVITY OF ARYL-BENZOYL-IMIDAZOLE (ABI) COMPOUNDS IN DRUG-
RESISTANT MELANOMA CELLS
[00455] P-glycoprotein (Pgp)-mediated drug efflux represents a major
mechanism for
cancer cells to prevent the build up of effective anticancer intracellular
drug concentrations. The
activity of the ABI compounds were compared against multidrug-resistant (MDR)
melanoma
cells (MDA-MB-435/LCCMDR1) and their parental nonresistant cancer cells (MDA-
MB-435).
Although MDA-MB-435 was originally designated as a breast cancer cell line, it
has been shown
definitively to originate from the M14 melanoma cell line. Compounds 12cla,
12th, 12cb, llcb,
and llfb together with other tubulin-targeting agents including colchicine,
paclitaxel, and
vinblastine were tested on both the MDR melanoma cell line and its parental
melanoma cell line
(Table 14A). Paclitaxel and vinblastine are clinically used anticancer drugs
known to target cell
tubulin. Although colchicine is not an FDA-approved drug for cancer treatment,
its prodrug,
ZD6126, is in clinical trial for solid tumors. Bortezomib is the first
therapeutic proteasome
inhibitor and was approved in 2003 by the FDA for use in multiple myeloma. ABT-
751 is known
to target the tubulin colchicine binding site. It is a promising drug
candidate in clinical trial for
children with relapsed or refractory neuroblastoma. Compounds 12da, 12M, 12cb,
llcb, llib
had much better resistance indices (3.0 for 12da, 0.9 for 12M, 1.3 for 12cb,
0.8 for llcb, 0.7 for
11M) than colchicine (65.8), paclitaxel (69.3), and vinblastine (27.5).
Although colchicine,
paclitaxel, and vinblastine showed excellent activity in nonresistant melanoma
cell lines (0.5-10
217
CA 3030689 2019-01-18
nM), these compounds were significantly less potent in the MDR melanoma cell
fine (277-658
nM). In contrast, 12cb, llcb, 111b had essentially equivalent potency on both
MDR (15 nM, 38
nM, 30 nM, 30 nM, 35 nM for 12da, 12Th, 12cb, llcb and 111b respectively) and
nonresistant
melanoma cell lines (5 nM, 41 nM, 24 nM, 38 nM, 50 nM for 12cla, 12th, 12cb,
llcb and lltb
respectively). Compound 12da was more active than paclitaxel and colchicine on
A375 and
WM-164 cells.
Table 14A. In vitro growth inhibitory effects of the ABI compounds in
comparison to other
anticancer drugs on multidrug-resistant melanoma cell line (MDR cell) and the
matching
sensitive parent cell line (Normal Melanoma cell).
IC50 SEM (nM) (n=3)
Compound Tubulin MDA- MDA-MB-
WM- Resistance
11) A375 B16-F1 binding MB- 435
164 index*
(pm) 435 /LCC6MDR1
12da 9 2 46 5 8 2 0.2 0.1 5 1 15 2 3.0
12Th 52 4 73 6 74 9 3.9 2.1 = 41 2 38 2 -- 0.9
12cb 31 5 63 7 28 3 3.4 1.5 24 2 30 4 1.3
llcb 36 5 71 8 43 6 ND 38 3 30 2 0.8
11Th 88 9 107 12 74 8 ND 50 6 35 3 0.7
Paclitaxel 12 3 17 2 1813 N/A 4 1 277+41 69.3
Vinblastine 1.1 0.2 4.7 0.7 0.6 0.1 ND 0.4 0.1 11+1 27.5
Colchicine 20 3 29 5 10 2 1.8 0.5 10 1 658 50 65.8
Bortezomib 8 1 24 2 8 1 ND ND ND ND
A B T-751 1111 108 2127 351 661 56 ND ND ND ND
*Resistance indexes were calculated by dividing IC50 values on multidrug-
resistant cell line
MDA-MB-435/LCC6MDR1 by IC50 values on the matching sensitive parental cell
line
MDA-MB-435. Abbreviations: N/A, value not available; ND, not determined.
20
218
=
CA 3030689 2019-01-18
Table 14B. Anticancer efficacy and colchicine site binding affinity of ABIs in
different cancer
and MDR cell lines with different resistance mechanisms. ABIs showed excellent
potency against
all tested melanoma cell lines including highly metastatic and multidrug
resistant cell lines. High
binding affinity of ABIs to the colchicine binding site in tubulin confirmed
their target inside
cells.
IC5,, SEM (nmol/L) (n=3)
12cb 12da 12fb Paclitaxel Vinblastine Colchicine ABT-751 SN-38
A375 31 5 9 2
52 4 12 3 1 0.1 20 3 685 108 ND
A375MA2 44 5 8+1 55 4 8 1 1 0.2 18+2 265 36 ND
B16-F1 63 7 46 5 73 6 17 2 5 1 29
5 2127 351 ND
WM-164 28 3 8 2 74 9 18 3 0.6 0.1 10 2 661 56 ND
MDR1
MDA-MB-
24+2 5+1 41 2 4+1 0.4 0.1 10 1 417 23 ND
MDA-MB-
30 -4 11 2 38 2 277 4 11 1 658 50 577 31
435/LCC6MD ND
RI (1) (2) (1) (69) (28) (66) (1)
0VCAR-8* 25+2 11 +1 45+2 10+0.2 2 0.1
12 1 785 Ii 2 0.2
NCl/ADR- 13 1 5 0.1 20 6 5109 170 570 84 737 -51 864 42 10 1
RES (0.5) (0.5) (0.4) (511) (285) (61) (1) (5)
MRP
HEK293 - 12 2 9 1 54 0.3 9 0.3 5 0.1 3 0.4
645 153 3 0.4
pcDNA3.1*
HEK293- 16 2 8 1 33 7 30 3 24 1 5 0.1 717 28 9 0.04
MRP1 (1) (0.9) (0.6) (3) (5) - (2) (1)
(3)
HEK293- 14+4 8 0.3 39 12 37 2 28 2 3
0.3 747 7 7 0.1
MRP2 (1) (0.9) (0.7) (4) (6) (1) (1) (2)
BCRP
HEK293- 17 1 8 1 23 3 50 1 25 1 5 0.1 653 72 123 28
482R2 (1) (9.9) (0.4) (6) (5) (2) (1)
(41)
Tubulin binding
(PM)* 3 I 0.2 0.1 4 1 N/A ND 2+1 3.1
ND
Notes: *: parental cell line to drug resistant cell subline; MDR1 were
overexpressed in MDA-MB-435/LCC6MDR I
and NCl/ADR-RES; MRP1, MRP2 and BCRP were overexpressed in HEK293-MRP I,
HEK293-MRP2, and
HEK293-482R2. The resistance indexes (numbers in the parenthesis) were
calculated by dividing IC50 values on the
resistant cell subline by that of the matching parental cell line. '1: IC50
for tubulin binding was calculated from
+4.
[311)colchicine competition-binding scintillation proximity assay. : binding
affinity reported in the literature for
ABT-751. Abbreviations: N/A, not applicable since they bind to tubulin at
different sites.
[00456] The results of Table 14A showed that cell line MDA-MB-
435/LCCMDR1 was
very resistant to colchicine, paclitaxel, and vinblastine. But the ABIs of
this invention showed
equal potency to the drug-resistant cell line and the sensitive parent cell
line. This result strongly
suggests that ABIs are not substrates for P-gp. Thus, they overcame the
multidrug resistance
found in MDA-MB-435/LCCMDR1 cells. The dose response curves are shown in
Figure 21 for
12M, 12da, and 12cb. Table 14B explores further the resistance mechanisms for
paclitaxel, SN-
219
CA 3030689 2019-01-18
38, vinblastine, and colchicine as compared to the ABIs 12cb, 12da, and 12fb.
MRP and BCRP
conferred moderate resistance to pacleitaxel (resistance indexes of 4 and 6,
respectively),
vinblastine (resistance indexes of 6 and 5, respectively), and BCRP conferred
significant
resistance to SN-38 (resistance index of 41). However, none of the ABIs were
susceptible to
MRP- or BCRP-mediated resistance (resistance indexes ranged from 0.4 to 1.0).
ABT-751, like
the ABIs, was not susceptible to MDR1, MRP, or BCRP.
EXAMPLE 16
IN VITRO MICROTUBULE POLYMERIZATION ASSAY
Materials and Methods
[00457] Bovine brain tubulin (0.4 mg) (Cytoskeleton, Denver, CO) was
mixed with 10 plYI
of the test compound and incubated in 110 p.1 of general tubulin buffer (80 mM
PIPES, 2.0 mM
MgCl2, 0.5 mM EGTA, and 1 mM GTP) at pH 6.9. The absorbance at 340 nm was
monitored
every 1 min for 15 min by the SYNERGY 4 Microplate Reader (Bio-Tek
Instruments, Winooski,
VT). The spectrophotometer was set at 37 C for tubulin polymerization.
Results
[004581 The inhibition of tublin polymerization by Aryl-Benzoyl-hnidazole
(ABI)
compounds was examined. Bovine brain tubulin (>97% pure) was incubated with
three potent
ABI compounds, 12cb, 12da, and 12db at a concentration of 10 p.M, to determine
the effect of
these ABI compounds on tubulin polymerization (Figure 22). Tubulin
polymerization was
completely inhibited by compound 12da, while ¨ 80% inhibition was observed
during incubation
with compounds 12cb and 12db.
[00459] This microtubule destabilization effect was similar to that of
colchicine and
vinblastine but was opposite to that of paclitaxel. The results not only
confirmed that ABIs can
directly interact with tubulin but also suggested that they may share the same
binding site with
colchicine (or vinblastine).
220
CA 3030689 2019-01-18
EXAMPLE 17
MELANOMA INHIBITION IN VITRO
Materials and Methods
[00460] B16-F1 melanoma cells were plated at a colony-forming density (2000
cells per
well on six-well plates) on top of 0.8% base agar. Cells were grown in 0.4%
agar together with
DMEM medium supplemented with fetal bovine serum and an antibiotic-antimycotic
solution at
37 C in an atmosphere of 95% air and 5% CO2. Cells were treated with
compounds 12da, 12cb
and 12Th at different concentrations (20, 100, and 500 nM). Compounds were
added to the media
from 1 mM DMSO stock solutions, and a corresponding dilution of DMSO was used
as control.
Cells were grown for 14 days. Plates were photographed, and the number of
colonies was
measured by Artek 880 Automated Colony Counter (Artek Systems Corporation,
Farmingdale,
NY).
Results
[00461] Four representative photos are shown in Figure 23. After 14 days of
incubation,
about 130 detectable colonies (diameter larger than 100 gm) were formed in
controls (no
treatment).
[00462] Compounds 120 and 12da effectively inhibited B16-F1 melanoma
colony
formation even at the lowest tested concentration, 20 nM (p<0.05 compared with
control). 12Th
showed effective inhibition at 100 nM. All three tested compounds showed
complete inhibition
of colony formation at 0.5 M, further proving ABIs antimelanoma efficacy.
EXAMPLE 18
=
IN VIVO ANTI-TUMOR ACTIVITY
Materials and Methods
[00463] Animals: Female C57/BL mice, age 4-6 weeks, were purchased
from Harlan
Laboratories (Harlan Laboratories Inc., Indianapolis, IN). The animal housing
met the
= Association for Assessment and Accreditation and Laboratory Animal Care
specifications. All of
the procedures were conducted in accordance with guidelines of our
Institutional Animal Care
and Use Committee.
221
CA 3030689 2019-01-18
{00464I In vivo evaluation of efficacy. Mouse melanoma B16-Fl cells
were prepared in
FBS-free DMEM medium (Cellgro Mediatech) at a concentration of 5 x 106 viable
cells/mL. The
cell suspension (100 L.) was injected subcutaneously in the right dorsal
flank of each mouse.
When tumor size reached about 100-150 mm3, about 7 days after cell
inoculation, all mice
bearing tumors were divided into control and treatment groups based on tumor
size (n = 5 per
group). Each group had similar average tumor size. Mice in control groups
(negative control)
were injected intraperitoneally with 50 pi- vehicle solution only or DTIC at
60 mg/kg (positive
control) once daily. Tumor volume was measured every 2 days with a traceable
electronic digital
caliper (Fisher Scientific, Inc., Pittsburgh, PA) and calculated using the
formula a x b2 x0.5,
where a and b represented the larger and smaller diameters, respectively.
Tumor volume was
expressed in cubic millimeters. Data were expressed as mean SE for each
group and plotted as
a function of time. Percentage tumor reduction at the conclusion of the
experiment (14 days after
starting treatment) was calculated with the formula 100-100 x [(T - To)/(C -
C0)], where T
represents mean tumor volume of a treated group on a specific day, T0
represents mean tumor
volume of the same group on the first day of treatment, C represents mean
tumor volume of a
control on a specific day, and Co represents mean tumor volume of the same
group on the first
day of treatment. Animal activity and average body weight of each group were
monitored during
the entire experiment period to assess compound toxicity. At the end of
treatment, all mice were
euthanized by CO2 followed by cervical dislocation, and tumors were harvested
for further
studies.
Results .
[00465] To evaluate efficacy of ABI analogs in vivo, we tested the
antitumor activity of
compound 12cb on mice melanoma B16-F1 xenograft. against DTIC, the gold
standard in
malignant melanoma treatment, was used as a positive control (Figure 24A).
Twenty female
C57/BL mice were divided into four groups: a vehicle control group, a DTIC (60
mg/kg)
treatment group, a 12cb (10 mg/kg) treatment group, and a 12cb (30 mg/kg)
treatment group.
Each mouse was injected with 0.5 million B16-F1 melanoma cells subcutaneously.
Seven days
after tumor inoculation, treatment started with each compound injected
intraperitoneally daily
(Figure 24). Tumor volume was significantly (p<0.05) reduced 47%, 51%, and 73%
for 12cb
(10 mg/kg), DTIC (60 mg/kg), and 12cb (30 mg/kg), respectively, after 14 days
of treatment. No
significant weight loss was observed in any of the treatment groups during the
experiment.
222
CA 3030689 2019-01-18
[00466] Two dose levels of 12113, 15 and 45 mg/kg, were chosen. DTIC at
60 mg/kg was
used as a positive control. B16-F1 melanoma allogyaft model on C57BL/6 mice
was first chosen
for study. After 13 days of treatment (Figure 24B), compound 121b inhibited
melanoma tumor
growth (TGI value) by 32% at 15 mg/kg and 82% at 45 mg/kg. Student's I test p
value of 12Th at
45 mg/kg compared with control was less than 0.001, indicating a significant
difference. The:
test p value of 121b at 15 mg/kg compared with control was 0.08, suggesting
that this dose was
not effective. Comparing 12Th at 45 mg/kg with DTIC at 60 mg/kg, which had a
TGI of 51%, the
t test p value was about 0.001, suggesting that 12Th had substantially better
activity than did
DTIC. For the control and 12Th 15 mg/kg treatment groups, average body weight
increased
slightly throughout the experiment period.
[00467] To further confirm ABIs' in vivo activity, A375 human Melanoma
xenograft model
on SHO mice was used, and 12fb at 25 mg/kg was tested. DTIC at 60 mg/kg was
used as a
positive control again. After 31 days of treatment (Figure 24C), 12Th
inhibited melanoma tumor
growth (TGI value) by 69%, whereas DTIC inhibited growth by 52%. The t test p
value of 12Th
.. treatment versus control was less than 0.001, suggesting that 12Th
significantly inhibited
melanoma tumor growth at 25 mg/kg. The t test p value of 12Th treatment versus
DTIC was less
than 0.05, suggesting again that 12Th had better activity than did DTIC.
Average body weight of
all groups increased slightly throughout the experiment period. Physical
activities for the mice
also looked normal, suggesting that 25 mg/kg was a well tolerated dose for SHO
mice.
EXAMPLE 19
BINDING TO COLCHICINE
Materials and Methods
[00468] Each test compound was prepared at 20 x concentration In G-PEM
buffer
(Cytoskeleton Inc., Denver, CO) followed by pipetting 10 L of test compound
into the 96-well
plates. Ten microliters of tritiated labeled colchicine (Perkin-Elmer,
Waltham, MA) was added to
each testing well. Subsequently, 180 1_, bead/tubulin (GE Healthcare Bio-
Sciences Corp.,
Piscataway, NJ) suspension was added into each well. The plate was incubated
for 45 min at 37
C before it was read by a Topcount NXT plate reader (Perkin-Elmer, Waltham,
MA).
Nonradiolabeled "cold" colchicine was included as a positive control and
paclitaxel as a negative
223
=
CA 3030689 2019-01-18
control because paclitaxel binds to a different site in tubulin and does not
compete for the
colchicine site binding. Data were processed using GraphPad Prism software.
Cell cycle analysis
[00469] Flow cytometry analysis was performed to study cell cycle phase
distribution.
A375 cells were cultured in 10-cm tissue culture dishes until the confluence
was about 80%, and
then cells were treated with 0, 10, 50, 200, and 1000 nM of colchicine, 12da,
12th and 12cb, for
24 h in growth media. Cellular DNA was stained with PBS containing 50 pg/mL
propidium
iodide and 100 pg/mL RNase A. The cell cycle was determined using a BD LSR-H
cytometer
(BD Biosciences, San Jose, CA) with 10,000 cells scored. Data were analyzed
and graphs were
prepared using the Modfit 2.0 program (Verity Software House, Topsham, ME).
Results
[00470] Three ligand binding sites in tubulin a/13-heterodimer have
been reported:
paclitaxel binding site, vinblastine binding site, and colchicinc binding
site. The binding affinity
of compound 12cb using 3H-labeled colchicine and a competitive binding
scintillation proximity
assay (SPA) was measured. The results confirmed the strong binding of 12cb
with a binding
affinity of 3.4 1.5 ttM (Figure 25A). Colchicine bound tubulin with an IC50
value of 1.8 0.5 pM
under these conditions. These results clearly indicated that ABI compounds
effectively inhibit '
tubulin polymerization.
[00471] The binding graph (Figure 25A) clearly shows that ABIs can
competitively bind
to the tubulin colchicine binding site. As the concentration of the three
tested compounds
increased from 0.03 KM to 100 p.M, increased tritiated colchicine was
competitively stripped
away from tubulin and emitted lower SPA counts. The negative control,
paclitaxel, gave only a
flat line, because theoretically it should not bind to the colchicine binding
site on tubulin. Second,
ABIs have relatively high binding affinity to the tubulin colchicine binding
site. GraphPad Prism
calculated IC50 values for binding showed that 12da has the highest binding
affinity. The binding
affinity was positively correlated to in vitro antimelanoma activity; the
higher the binding
affinity, the higher the antimelanoma activity.
[00472] ABIs demonstrated that they arrest cells by cell cycle analysis
in the G2/M phase
as indication that they target tubulin. Compounds 12da, 12113 and 12th were
tested together with
colchicine as a positive control on A375 cells (Figure 25B). Four different
concentrations - 10,
224
CA 3030689 2019-01-18
50, 200, and 1000 nM - of each compound were chosen to show the dose effect
(Figure 25C and
25D). For controls (no treatment) without interference, about 16% of A375
cells were distributed
in the G2/M phase. For the colchicine treatment group, as concentration
increased from 10 n.M to
50 nM, the percentage of cells distributed in the G2/M phase increased from
14% to 85%. ABIs
had similar results for A375 cells, in arresting them in the G2/M phase in a
dose-dependent
manner. The potency of the different concentrations in arresting cells in the
G2/M phase
positively correlated with in vitro activity.
EXAMPLE 20
IN VITRO AND IN VIVO PHARMACOLOGY OF COMPOUNDS 17va, 12fa, AND 55
Materials and Methods
[004731 Cell culture and cytotoxicity assay of prostate cancer. All
prostate cancer cell
lines (LNCaP, PC-3, and DU145, PPC-1) were obtained from ATCC (American Type
Culture
Collection, Manassas, VA, USA). Human PC-3_TxR, was resistant to paclitaxel
and used a MDR
model compared with PC-3. Cell culture supplies were purchased from Cellgro
Mediatech
(Herndon, VA, USA). All cell lines were used to test the antiproliferative
activity of compounds
17ya, 12fa, and 55 by sulforhodamine B (SRB) assay. All cancer cell lines were
maintained in
RPMI 1640 media with 2 mM glutamine and 10% fetal bovine serum (FBS).
[00474] In vitro microtubule polymerization assay. Porcine brain
tubulin (0.4 mg)
(Cytoskeleton, Denver, CO) was mixed with 1 and 5 xM of the test compound or
vehicle
(DMSO) and incubated in 100 tIL of buffer (80 mM PIPES, 2.0 mM MgCl2, 0.5 niM
EGTA, pH
6.9 and 1 mM GTP). The absorbance at 340 nm wavelength was monitored every min
for 15 min
(SYNERGY 4 Microplate Reader, Bio-Tek Instruments, Winooski, VT). The
spectrophotometer
was maintained at 37 C for tubulin polymerization.
= [00475] Metabolic incubations. Metabolic stability studies
were conducted by incubating
0.5 04 of test compounds in a total reaction volume of I mL containing 1 mg/mL
microsomal
protein in reaction buffer [0.2 M of phosphate buffer solution (pH 7.4), 1.3
mM NAM), 3.3 mM
glucose-6-phosphate, and 0.4 U/mL glucose-6-phosphate dehydrogenase] at 37 C
in a shaking
water bath. The NADPH regenerating system (solution A and B) was obtained from
BD
225
CA 3030689 2019-01-18
Biosciences (Bedford, MA). For glucuronidation studies, 2 mM UDP-glucuronic
acid (Sigma, St.
Louis, MO) cofactor in deionized water was incubated with 8 rnM MgC12, 25 p_g
of alamethicin
(Sigma, St. Louis, MO) in deionized water, and NADPH regenerating solutions
(BD Biosciences,
Bedford, MA) as described previously. The total DMSO concentration in the
reaction solution
was approximately 0.5% (v/v). Aliquots (100 4) from the reaction mixtures used
to determine
metabolic stability were sampled at 5, 10, 20, 30, 60, and 90 min.
Acetonitrile (150 AL)
containing 200 nM of the internal standard was added to quench the reaction
and to precipitate =
the proteins. Samples were then centrifuged at 4,000g for 30 min at RT, and
the supernatant was
analyzed directly by LC-MS/MS.
[00476] Analytical method. Sample solution (10 m.L) was injected into an
Agilent series
HPLC system (Agilent 1100 Series Agilent 1100 Chemstation, Agilent Technology
Co, Ltd). All
analytes were separated on a narrow-bore C18 column (Alltech Alltima HP,
2.1x100 mm, 3 um,
Fisher, Fair Lawn, NJ). Two gradient modes were used. For metabolic stability
studies, gradient
mode was used to achieve the separation of analytes using mixtures of mobile
phase A
[ACN/H20 (5%/95%, v/v) containing 0.1% formic acid] and mobile phase B [ACN/1-
I20
(95%/5%, v/v) containing 0.1% formic acid] at a flow rate of 300 glimin.
Mobile phase A was
used at 10% from 0 to 1 min followed by a linearly programmed gradient to 100%
of mobile -
phase B within 4 mm, 100% of mobile phase B was maintained for 0.5 min before
a quick ramp
to 10% mobile, phase A. Mobile phase A was continued for another 10 min
towards the end of
.. analysis.
[00477] A
triple-quadruple mass spectrometer, API Qtrap 4000111 (Applied
Biosystems/MDS SCIEX, Concord, Ontario, Canada), operating with a
TurbolonSpray source
was used. The spraying needle voltage was set at 5 kV for positive mode.
Curtain gas was set at
10; Gas 1 and gas 2 were set 50. Collision-Assisted-Dissociation (CAD) gas at
medium and the
source heater probe temperature at 500 C. Multiple reaction monitoring (MRIM)
mode, scanning
m/z 378 ¨4 210 (17ya), m/z 373 --4 205 (12fa), rn/z 410 242
(55) and irdz 309 171 (internal
standard), was used to obtain the most sensitive signals. Data acquisition and
quantitative
processing were accomplished using Analyst software, Ver. 1.4.1 (Applied
Biosystems).
[00478] Aqueous
solubility. The solubility of drugs was determined by Multiscreen
Solubility Filter Plate (Millipore Corporate, Billerica, MA) coupled with LC-
MS/MS. Briefly,
226
CA 3030689 2019-01-18
198 1.1.L, of phosphate buffered saline (PBS) buffer (pH 7.4) was loaded into
96-well plate, and 2
lit of 10 mM test compounds (in DMSO) was dispensed and mixed with gentle
shaking (200-
300 rpm) for 1.5 hours at RI (N = 3). The plate was centrifuged at 800g for 10
min, and the
filtrate was used to determine its concentration and solubility of test
compound by LC-MS/MS as
described previously.
[00479] Pharmacokinetie study. Male ICR mice (n = 3 per group) 6 to 8
weeks of age
were purchased from Harlan Inc., and used to examine the pharmacokinetics (PK)
of 17ya, 12fa,
and 55. All compounds (10 mg/kg) were dissolved in DMSO/PEG300 (1/9) and
administered by
a single intravenously (i.v.) injection (50 tit) into the tail vein. Blood
samples were collected at
5, 15, and 30 min, 1, 1.5, 2, 3,4, 8, 12, and 24 h after i.v. administration.
Mice were given (p.o,)
by oral gavage at 20 mg/kg (in Tween80/DMSO/H20, 2/2/6) of each test compound
to evaluate
their oral bioavailability. Blood samples were collected at 0.5, 1, 1.5, 2,
3,4, 8, 12, and 24 h after
p.o. administration.
[00480] Female Sprague-Dawley rats (n = 3; 254 4 g) were purchased
from Harlan Inc.
(Indianapolis, IN). Rat thoracic jugular vein catheters were purchased from
Braintree Scientific
Inc. (Braintree, MA). On arrival at the animal facility, the animals were
acclimated for 3 days in a
temperature-controlled room (20-22 C) with a 12 h light/dark cycle before any
treatment.
Compounds 17ya, 12fa, and 55 were administered i.v. into the thoracic jugular
vein at a dose of 5
mg/kg (in DMSO/PEG300, 1/9). An equal volume of heliarinized saline was
injected to replace
the removed blood, and blood samples (250 }iL) were collected via the jugular
vein catheter at
10, 20, 30 min, and 1, 2, 4, 8, 12, 24 h. Rats were given (p.o.) by oral
gavage at 10 mg/kg (in
Tween80/DMSO/H20, 2/2/6) of each test compound to evaluate their oral
bioavailability. All
blood samples (250 pl.-) after oral administration were collected via the
jugular vein catheter at
30, 60, 90 min, 120 min, 150 min, 180 min, 210 min, 240 min, and 8, 12, 24 h.
Heparinized
syringes and vials were prepared prior to blood collection. Plasma samples
were prepared by
centrifuging the blood samples at 8,000g for 5 min. All plasma samples were
stored immediately
at -80 C until analyzed.
[00481] Analytes were extracted from 100 pL of plasma with 200 u.L of
acetonitrile
containing 200 nM the internal standard. The samples were thoroughly mixed,
centrifuged, and
the organic extract was transferred to autosampler for LC-MS/MS analysis.
227
CA 3030689 2019-01-18
[00482] PC-3_TxR xenograft studies. PC-3_TxR cells (10x102 per mL) were
prepared in
RPMI1640 growth media containing 10% FBS, and mixed with Matrigel (BD
Biosciences, San
Jose, CA) at 1:1 ratio. Tumors were established by injecting 100 p.L of the
mixture (5x106 cells
per animal) subcutaneously (s.c.) into the flank of 6-8-week-old male athymic
nude mice. Length
and width of tumors were measured and the tumor volume (mm3) was calculated by
the formula,
11/6 x L x W2, where length (L) and width (W) were determined in mm. When the
tiimor volumes
reached 300 mm3, the animals bearing PC-3_TxR tumors were treated with vehicle
[Tween80/DMSO/H20 (2/2/6)], or 17ya (10 mg/kg) orally. The dosing schedule was
3 times a
week for four weeks,
Results
[00483] 17a and 55 exhibit broad cytotoxicity in cells, including
multidrug-resistant
cells. The ability of 17ya and 55 to inhibit the growth of cancer cell lines
was evaluated using
SRB assay (Table 15). Both compounds inhibited the growth of Several human
cancer cell lines,
including five prostate and one glioma cancer cell lines, with IC50 values in
the low nanomolar
range. 17ya exhibited L7-4.3 fold higher potency than 55 in these cell lines.
Paclitaxel-resistant
PC-3 (PC-3/TxR) cell line that over-expresses P-glycoprotein (P-gp), was used
to study the effect
of drug resistance on 17ya and 55 and to compare against its parent, PC-3 cell
line. The IC50
values of docetaxel were 1.2 0.1 nM and 17.7 0.7 nM in PC-3 and PC-3/TxR
cells,
respectively. 17ya and 55 were both equipotent against parent PC-3 and PC-
3/TxR, whereas
paclitaxel and docetaxel exhibited relative resistance of 85- and 15-fold,
respectively. These data
indicate that both 17ya and 55 circumvent P-gp-mediated drug resistance.
Table 15. Cytotoxicity data of 17ya and 55.
228
CA 3030689 2019-01-18
Cell line Type Cytotoxicity [ICso values, mean SD nIVI]
17ya 55 Paclitaxel
s
.1) >44 0 0 =,:y
0
'143
PC-3 Prostate 5.2 0.2 16 1.5 0.6 0.05
PC-3/TxR Prostate 2.1 - 0.1 (0.4) 6.7 0.5 (0.4) 51
2.3 (85)
LNCaP Prostate 12 0.1 27 0.6 1.7 0.2
Du-145 Prostate 17 + 0.2 38 0.6 5.1 0.1
PPC-1 Prostate 21 + 0.1 36 0.4 2.3 0.8
U87MG Glioma 10 1.6 22 3.0 NR
1050 values (mean SD) were determined after 96 h treatment (N = 3).
Paclitaxel was used as a
positive control. Data in parentheses indicated resistance factor when
compared IC50 values in
PC-3 and PC-3/TxR. NR, Not Reported.
=
[00484) 17ya and 55 bind to colchicine-binding site on tubulin, inhibit
tubulin
polymerization, and induce cell apoptosis (Figure 26). A competitive mass
binding assay was
developed to study the interaction of small molecule inhibitors with tubulin.
In this study, varying
concentrations of 17ya or 55 were used to compete with colchicine-tubulin
binding. Both
compounds competed effectively with colchicine for tubulin binding (Figure
26A); however,
their competitive binding curves deviated substantially from zero at higher
concentrations when
compared to podophylltoxin, a known potent colchicine-site binding ligand.
This suggests that
both 17ya and 55 exhibited less affinity than podophylltoxin or they partially
bind to the
colchicine-binding site. Vinblastine, the negative control, did not inhibit
the colchicine-tubulin
binding, successfully demonstrating the specificity of this competitive mass
binding assay
[00485] Porcine brain tubulin (>97% pure) was incubated with 17ya or 55
(5 1..tM) to test
their effect on tubulin polymerization (Figure 2613). 17ya and 55 inhibited
tubulin
polymerization by 47% and 40% at 15 min, respectively. Colchicine at 5 i.tM
was used as a
positive control and inhibited tubulin polymerization by 32%. These data
suggest that both 17ya
and 55 have slightly greater inhibition of tubulin polymerization than
colchicine. Therefore, the
229
CA 3030689 2019-01-18
molecular mechanism of these compounds is binding to the colchicine-binding
site, inhibiting
tubulin polymerization, and inducing cytotoxicity.
[00486] PC-3 and PC-3/TxR cells were exposed to 0.8 to 600 nmol/L of
17ya, SS, or
docetaxel for 24 h. The levels of DNA-histone complexes were used to represent
cell apoptosis.
Both 17ya and 55 were equally potent to induce cell apoptosis in PC-3 (Figure
26C) and PC-
3/TxR (Figure 26D) in 24 h. Though, docetaxel was highly potent to induce
apoptosis of PC-3
cells, it was weaker in PC-3/TxR cells due to over-expression of P-gp.
[00487] 17ya and 55 exhibited favorable drug-like properties. Drug-like
properties,
such as metabolic stability, permeability, aqueous solubility, and drug-drug
interactions, were
examined for 17ya and 55 (Table 16A). 17ya exhibited greater metabolic
stability, and aqueous
solubility than 55. Both chemicals exhibited more than adequate permeability
values, suggesting
their potential to be orally used. In addition, both 17ya and 55 showed high
IC50 values in
micromolar range on CYP enzyme inhibition assays, indicating that both
compounds may avoid
drug-drug interactions through main CYP liver enzymes. Overall, both compounds
exhibited
favorable drug-like properties.
Table 16A. Drug-like properties of compound 17a and 55. Metabolic stability,
permeability,
solubility, and potential drug-drug interactions were evaluated. Each value
represents the mean
from duplicate studies.
=
positive controls
Measurment Units 17ya SS
(mean)
Metabolic stability
half-life in human liver microsomes min > 60 28 Veraparnil
(12)
Permeabilityp9C =
P in CaC0-2 assay 104 cm/s 36 99
Propranolol
(19)
lh
Aqueous solubility tqlmL 75 19
(1.1)
Drug-drug interactions
value in Cyp3A4 Ketoconazoie
5.5
(substrate: Testosterone) (0.02)
IC50 value in Cyp206 Ouinindine
uM (substrate: Oextrornethorphart) > 50 34 (0.1)
IC,, value in Cyp2C19 M 66 3 Ticlopidine
N 5
(substrate: (S)-meph . enytoin) (0.37)
IC,, value in Cyp2C9 17 4 9 Sulfaphenazole
, .
(substrate: Diclotenac) (0.5)
IC5c value in Cyp1A2 ft4 92 8 1 Furafylline
il
(substrate: Phenacetin) . (2.2)
230
CA 3030689 2019-01-18
r)
P-74392-PC1
co
0 Table 16B. Summary of drug-like and pharmacokinetic properties of
17ya, 12fa, 55, and lh.
0
0,
cc, = , 1 7ya 1_ 12fa
i 55 1h
l0 0
0 0
IQ 0.,
ii.4 . a
0 HN 110 HN -"N
ao 0-2 ir :,.. Ai. 0.
0- ,c7- 0- Ht,s_, 0,. d-N
lir to
1 o, o,
0 HN
N
CO
Molecular weight 377 372
409 355
ICso in PC3 (nM) nM 10 35
28 = 21
Half-life in HLM (Phase!) min -80 ' 44
30 17
Half-life in HLM (Phase 1+11) min -90 NA
43 17
Solubility ilg/mL ' >75 12
19 1 .
RatPK_IV5mgk_CI m Umin/kg 16 '.
', ' ' 7.7 (2.5mpk)
RatPK_IV5mgk_V Ukg . , 1.9 -
' = ' 4.9 (2.5mpk)
RatPK_P010mgk_Cmax ng/mL , 1109
212
RatPK_P010mgk_AUC min*Ag/mL 218 .
= . 37
RatPK_Bioavailability 0/oF 35
3.3
,
.
MousePK_IV1Orngk_CI mL/min/kg . 61
130
. .
.e. = -
MousePK_IV10mgk_V Ukg 4
4.9
=
MousePK_P020mgk_Cmax ng/mL , 2592 .
NA
MousePK_P020mgk_AUC min*gg /mL ' = 201
NA
MousePK_Bioavailability %F .. = 62
NA
,
231
[00488] As shown in Table 16B, 17ya had a half-life of 80 min by phase
I reaction,
suggesting that 17ya was stable in phase I metabolic processes. The half-life
(90 min) in the
presence of UDP-glucuronic acid was similar to that observed in its absence.
These data
suggested that 17ya is stable in human liver microsomes, and it was hoped that
low clearance and
long half-life will be obtained in human. On the other hand, 55 exhibited 30
and 43 min as half
lives when it was in the presence and absence of UDP-glucuronic acid,
respectively. Compound
12fa shows the half-life with 44 in phase I. These data suggested that all
three compounds
showed acceptable stability in human liver microsomes, and 17ya is more stable
than 12fa and
55. When investigating their metabolism, it was found that 12fa and 55
exhibited higher levels of
ketone-reduction (data not shown), suggesting that 12fa and 55 are more labile
than 17ya.
[60489] Compound 17ya exhibited great aqueous solubility, 12fa and 55
showed
acceptable solubility.
Compound 17ya contained an imidazole ring, and this ring improved aqueous
solubility,
resulting in > 75 pt/mL aqueous solubility (Table 16A). Compounds 12fa and 55
exhibited less
aqueous solubility, and exhibited 12 and 19 gg/mL, respectively. Overall, 17ya
demonstrated a
great aqueous solubility, and 12fa and 55 showed acceptable aqueous
solubility, and much
improved over lb. The greater solubility of 121a translated into much improved
oral
bioavailability compared to lh (35% vs. 3.3% in rat). Similarly for 17ya and
55, aqueous
solubility correlated with much improved oral bioavailability as discussed
infra (Table 17).
[00490] Pharmacokinetic studies of 17ya and 55 in mice, rats and dogs. The
pharmacokinetic parameters of 17ya and 55 given in a single (i.v. or p.o.)
dose in ICR mice,
Sprague-Dawley rats, and beagle dogs are summarized in Table 17. 17ya
exhibited low clearance
in mice arid rats, suggesting that 17ya exhibited metabolic stability, and
minimal first-pass
metabolism in these species. In addition, 17ya had moderate volume of
distribution in mice and
rats, indicating that it may properly distribute into tissues, including
tumors. Unlike in mice and
. rats, surprisingly, the total clearance of 17ya in dogs was high. Two
abundant metabolites in dog
plasma, a hydroxylated metabolite and an unknown metabolite with +34 m/z of
the parent (data
not shown), were consistent with those found in dog liver microsomes. In
summary, higher
clearance and lower oral exposure was obtained for 17ya compared to 55 in
dogs, but not in mice
and rats. In addition, 17ya exhibited abundant metabolites only in dog liver
microsomes, but not
in mouse, rat or human liver microsomes (data not shown). 17ya showed
acceptable 21%, 36%,
232
CA 3030689 2019-01-18
= and 50% oral bioavailability in rats, mice, and dogs, respectively.
Meanwhile, 55 had low
clearance in rats, and moderate clearance in mice and dogs. Similar to 17ya,
55 exhibited
moderate volume of distribution in these species. 55 had constant oral
bioavailability rates among
three species (24%-36%). These properties indicate that both 17ya and 55 are
potential orally
available tubulin inhibitors.
[00491] Table 17. Phanmacokinetic studies of compounds 17ya and 55 in
mice, rats, and
dogs.
17ya 55
IV PO IV Po
Mouse PK (N=3)
Dose, mg/kg 10 20 10 20
Clearance, MUmin/kg 19 NR 40 NR
Vss, Ukg 2.9 NR 1.3 NR
t, min 101 339 46 125
AUC, min'Itg/mL 540 384 . 249 171
C,, ng/mL 4800 1560 7739 1253
36% 34%
Rat PK (N=3)
Dose, mg/kg 5 ' 10 5 10
Clearance, mUminIkg 9.5 2.3 NR 10 1.4 NR=
Vss, Ukg 1.8 0.2 NR 1.0 0.1 NR
t,,2, min 139 24 206 12 73 5.0 350 t214
-
AUC, min*ug/mL 553 143 233 134 509 73 246 i
163
Cõ ng/mL 3672 519 999 445 4609 55 757
520
21% 24%
Dog PK (N=4)
Dose, mg/kg 2 5 2 5
Clearance, mUmin/kg 109 29 NR 15 3.2 NR
'
Vss, Ukg 94 95 NR 0.9 0.2 NR
11,2, min 2757 1573 1695 439 82 15 191
9.0
AUC, min'rng/mL 18.5 4.7 23.1 11.3 141 30 128 154
C,,,,,u, ng/mL 400 118 210 133 2552 576 862
1010 .
50% 36%
[00492] 17ya and 55 inhibit paclitaxel resistant prostate (PC-3/TxR)
xenografts
growth. PC-3 (Figure 27A) and paclitaxel-resistant prostate cancer (PC-3/TxR)
(Figure 27B)
cells were inoculated in nude mice and the tumor volumes were allowed to reach
about 150-300
mm3. Docetaxel (10 or 20 mg/kg), which is in clinic for prostate cancer, was
used to evaluate its
effectiveness in models of P-gp-mediated drug resistance in vivo. PC-3/TxR
tumor was found to
be fast-growing and the volume reached 1500-2500 mm3 at the termination of the
study. Though
233
CA 3030 6 8 9 2 0 1 9 ¨0 1-1 8
and 20 mg/kg intravenously administered docetaxel exhibited a dose response in
both models
(Figures 27A and 27B), the tumor growth inhibition (TOO effect decreased from
84% TGI in
PC-3 tumors to 14% TGI in PC-3/TxR tumors when intravenously dosed at 10 mg/kg
(Table 18).
In addition, at the higher dose (20 mg/kg), docetaxel elicited partial
regression (>100% TGI) of
5 PC-3 tumors, but barely 56% TGI in PC-3/TxR tumors. The effectiveness of
docetaxel in PC-
3/TxR tumors was dramatically decreased when compared to that in PC-3 tumors,
suggesting that
the efficacy was impaired by P-gp-mediated drug resistance, and these results
are in very good
agreement with our in vitro cytotoxicity or apoptosis data. In contrast to the
lack of efficacy of
docetaxel in PC-3/TxR tumors, orally administered 17ya (6.7 mg/kg)
demonstrated more than
10 100% TGI without an effect on their body weights (Figure 27B and Table
18). In addition, 2 out
of 4 nude mice bearing PC-3/TxR tumors were tumor free on day 19 (data not
shown),
The PC-3/TxR xenograft model was further utilized to evaluate efficacies of
17ya (in other
dosing schedules) and 55. The maximal tolerated dose (body weight loss > 20%)
of 17ya was
found to be 10 mg/kg, when orally dosed once daily for four days; or at 3.3
mg/kg twice a day
(b.i.d.) for five days (data not shown). As shown in Figure 27C, 3.3 mg/kg of
17ya was dosed
b.i.d. for first consecutive four days in the first week, and the schedule was
then changed to once
daily between weeks 2 and 4. The result shows that partial regression was
obtained during day 4-
19, and the TGI was 97%; and one of the seven mice was tumor free on day 26.
Higher dose (10
mg/kg) with lower dosing frequency (q2d) of 17ya (Figure 27D) elicited partial
regression
during days 13 to 29. These data suggest that regimens with optimized doses
and dosing
schedules will facilitate 17ya to successfully inhibit PC-3/TxR tumors. 55,
was orally
administered to nude mice with 10 or 30 mg/kg b.i.d., and five times a week
between weeks 1
and 4. As shown in Fig 27C, the inhibition profiles exhibit a dose-response in
PC-3/TxR tumor.
The TGI value was 59% for the treatment group with a lower dose (10 mg/kg).
Moreover, the
higher dose (30 mg/kg) started to show partial regression (>100% TGI) from day
19 to the
termination of the study (day 26). Some mice in the vehicle group lost body
weight at the
endpoint, in part, due to cancer cachexia. On the contrary, mice treated with
17ya (3.3 mg/kg) or
55 (30 mg/kg) were gaining weight (Table 18), suggesting that these optimized
doses of 17ya or
55 may be well-tolerated and were preventive of cancer cachexia.
Table 18. Antitumor activity of compounds 17ya and 55 versus concomitantly
evaluated
docetaxel in vivo.
234
CA 3030689 2019-01-18
Dosing End Number Body weight (9) Tumor size (mm)
TGI (1S)
Schedule point End/Start Start End Start End
PC3 xenograft
Vehicle_IV day land 9 day 19 6/8 30 2 32 t 4 271 t
83 875 292 ¨
Docetaxel IV_10n174/ day land 9 day 19 5/5 29 5 2 24 t 2 247 2
49 341 101 84
0ocetaxer_IV_20mp6 day land 9 day 19 5/5 28 t 3 24 i 3 243268
172262 > 100 =
PC-31T2R xenograll
Vehicle IV day land 9 day 19 5/5 33 2 1 26 t 5 171 1
57 2061s 858 ¨
Docetaxel_IV_10mpk day land 9 day 19 4/4 31 t 2 2552
143 1 20 1774 5 183 14
Docetaxel_IV_20mp6 day land 9 day 19 4/4 3011 252.4 170 t 86
999t905 56
1 lya_P0_6.7mpk qd A 5/w day 19 4/4 33 t 3 34 t 3 172 I
69 126 100 .. 100
Vehlde_PO bid A 51w day 26 6/7 30 t 2 25 2 ..
156 30 2591 1423 .. ¨
55_P0_10mpk bid > 51w day 26 717 29 2 26 t 3 ..
143 t 44 1152 t 433 .. 59
55_P0_30mp6 bid x 5/w day 26 717 2913 3012 134234
101:19 > 100
17ya_PO 3_3Mple qd , 5/W day 26 7/7 29 2 3012 139244
2141172 97
Vehide PO q2d , 3/w day 29 5/5 24 t 2 21 21 299
40 1521 / 580 ¨
17ya P-0 lOmpk q2d a 3Iw day 29 5/5 24 2 2822 ..
294 156 237 1 103 .. a 100
Dosing schedule: qd x 5/w = one administration given on five consecutive days
per week; b.i.d.
x 5/w = two administrations given on five consecutive days per week; or q2d x
3/w = every other
day administration or three times a week.
a Dose schedule was two administrations given on four consecutive days of the
first week, and _
dose schedule was changed (because of toxicity) to one administration given on
five consecutive
days per week for the second to fourth week.
[00493] Brain penetration of 17ya and 55 in nude mice. Whole brain
concentrations in
nude mice at 1 h and 4 h after oral administration of 20 mg/kg 17ya or 55 were
determined
(Table 19). The ratios of brain to plasma concentrations were determined and
compared to
docetaxel in the nude mice. 55 exhibited greater brain penetration than 17ya
and docetaxel. 17ya
only exhibited slightly greater brain/plasma concentration ratios than
docetaxel at both 1 and 4 h.
The brain concentrations of 55 reached 14 to 19% of plasma concentrations at 1
h and 4 h,
respectively, showing a 3.2-fold higher brain/plasma ratio at both I h and 4 h
compared to
docetaxel. These data suggest that 55 exhibited potentially favorable
properties to treat glioma,
since it has greater brain penetration and high potency (22 nM, Table 15) in
glioma cells.
=
[004941 Table 19. Brain-Blood Barrier (BBB) studies of compounds 17ya and
55. Brain
and plasma concentrations were determined in nude mice at 1 and 4 h after
administration of
docetaxel (1P, 10 mpk), 17ya (PO, 20 mpk), and 55 (PO, 20 mpk). Each value
represents the
. mean SD from 3 nude mice.
235
CA 3030689 2019-01-18
Docetaxel 17ya 55
Measurment 1hr 4hr 1 hr 4hr 1 hr 4hr
Brain (ng/mL) 33 14 20 9 124 108 49 t 32 180 44
73 18
Plasma (ng/mL) 766 1 92 345 94 2058 1252 570 438
1609 867 380 t 32
Brain/plasma (%) 4.4 2.0 6.0 2.9 5.4 1.9 8.9 1.7
14 7.9 19 3.1
= EXAMPLE 21
PHARMACOKLNETICS OF COMPOUNDS OF THIS INVENTION
- Table 20.
Compound Half life in Half life in Half life in Half
life in Half life in
ID Human liver Mouse liver Rat liver Dog liver Monkey
microsome microsome microsome microsome liver
(min) (min) (min) (min) microsome
(min)
lb 17 <5 31 19 <5
¨
2e-cis 35
21 - - 32
2k 10 - 9 32 16 <5
21 20 11 49 30 8
_
6a 32 3.43 12 13 16
6b 40 ' 10 9 30 13
, _
6c 47 13 14 29 9
7d 24 37 42 29 15
12da 23 8 28 17 '
12fa 56 23 46 26
. _ .
12fb 37
-
12dab 21 <5 12 46
--
236
CA 3030689 2019-01-18
=
EXAMPLE 22
BIOLOGICAL ACTIVITY OF 4-SUBSTITUTED METHOXYBENZOYL-ARYL
THIAZOLE (SMART) COMPOUNDS lh. 2k, and 21: ACTIVE MICROTUBULE
INHIBITORS
Materials and Methods
[00495] In vitro microtubule polymerization assay. Bovine brain tubulin
(0.4 mg)
(Cytoskeleton, Denver, CO) was mixed with 10 pM of the test compound or
vehicle (DMSO)
and incubated in 100 I of buffer (80 mM PIPES, 2.0 mM MgCl2, 0.5 mM EGTA, pH
6.9 and 1
mM GTP). The absorbance at 340 nm wavelength was monitored every min for 15
min
(SYNERGY 4 Microplate Reader, Bio-Tek Instruments, Winooski, VT). The
spectrophotometer
was maintained at 37 C for tubulin polymerization.
[00496] ' MS competition binding assay. Colchicine, vinblastine, and
paclitaxel (1.2 pM
for each) were incubated with tubulin (1.2 mg/mL) in the incubation buffer (80
mM PIPES, 2.0
mM MgCl2, 0.5 mM EGTA, pH 6.9) at 37 C for 1 hr. lh (0.5-125 M) was examined
to
individually compete with colchicine-, vinblastine-, and paclitaxel-tubulin
binding. The free-form
ligands were separated from tubulin or microtubule using an ultrafiltration
method
(microconcentrator) (Microcon, Bedford, MA) with a molecular cutoff size of
30k Da.
Colchicine, vinblastine and paclitaxel were determined by LCMS/ MS method. The
ability of lh
to inhibit the binding of ligands was expressed as a percentage of control
binding in the absence
of any competitor. Each reaction was run in triplicate.
[00497] Cell culture and cytotoxicity assay of prostate and melanoma
cancer. All
prostate and melanoma cell lines were obtained from ATCC (American Type
Culture Collection,
Manassas, VA, USA), while cell culture supplies were purchased from Cellgro
Mediatech(Hemdon, VA, USA). The antiproliferative activity of the compounds
was examined
in four human prostate cancer cell lines (LNCaP, DU 145, PC-3, and PPC-1) and
two human
melanoma cell lines (A375 and WM-164). Human ovarian cell line OVCAR-8 and its
resistant
cell line that over-expresses P-gp, NCl/ADR-RES, were used as MDR models. Both
ovarian cell
237
CA 3030689 2019-01-18
lines were obtained from National Cancer Institutes (NCI). All prostate cancer
cell lines were
cultured with 10% fetal bovine serum (PBS).
[00498] Cell cycle analysis. Flow cytometry was performed to study the
effects of the
compounds on cell cycle distribution. PC-3 and A375 cells were treated in
growth media with the
indicated concentrations of compounds lh, 2k, 21 for 24 h. Cellular DNA was
stained with 100
pg/mL propidium iodide and 100 p.g/mL RNase A in PBS and flow cytometry was
performed to
determine the cell cycle distribution of the cells.
[00499] Apoptosis detection by ELISA. Quantification of the enrichment
of mono- and
oligonucleosomes in the cytoplasm was used to determine the ability of the
compounds to induce
apoptosis (cell death detection ELISA PLUS, Roche, Germany) following the
manufacturer's
=
instructions.
[00500] Pharmacokinetic study. Male ICR mice (n = 3 or 4 per group) 6
to 8 weeks of
age were purchased from Harlan Inc., and used to examine the pharmacokinetics
(PK) of the
compounds. lh, 2k, 21(15 mg/kg) were dissolved in PEG300/DMS0 (1/4) and
administered by a
single i.v. injection into the tail vein. Blood samples were collected at 2,
5, 15, and 30 min, 1, 2,
4, 8, 16, and 24 hr after administration. Male Sprague-Dawley rats (n = 4;
254, 4 g) were
purchased from Harlan Inc. (Indianapolis, 1N). lh, 2k, were administered
intravenously into the
, jugular venous catheters at 2.5 mg/kg (in DMSO/PEG300, 1/4). Blood samples
(250 L) were
collected at 10, 20, 30 min, and 1, 2, 4, 8, 12, 24, 48 h. A protein
precipitation method was used
for sample preparation. An aliquot (200 pL) of acetonitrile (ACN) was added to
100 AL of
plasma and then was thoroughly vortexed for 15 s. After centrifugation, the
supernatant was
analyzed by liquid chromatography tandem mass spectrometry (LC-MS/MS). The PK
parameters
were determined using Non compartment analysis (WinNonlin, Pharsight
Corporation, Mountain
View, CA).
[00501] PC-3 and A375 tumor xenograft studies. PC-3 and A375 cells (5x107
per mL)
were prepared in phenol red-free growth media containing 10% PBS, and mixed
with Matrigel
(BD Biosciences, San Jose, CA) at 1:1 ratio. Tumors were established by
injecting 100 pL of the
mixture (2.5x106 cells per animal) subcutaneously (s.c.) into the flank of 6-8-
week-old male
athymic nude mice. Length and width of tumors were measured and the tumor
volume (mm3)
was calculated by the formula, 7t/6 xL xW2, where length (L) and width (W)
were determined in
mm. When the tumor volumes reached 150 mm3, the animals bearing PC-3 tumors
were treated
238
CA 3030689 2019-01-18
with vehicle [Captex200fTween80 (1/4)], lh (5 and 15 mg/kg), 2k (5 and 15
mg/kg) and 21(50
mg/kg) intraperitorally for 21 days. Vinblastine (0.5 mg/kg) was used as the
positive control and
dosed q2d with vehicle [DMSO/PEG300 (I/9)]. On the other hand, A375 tumor
bearing mice
were treated for 34 days with vehicle [Captex200/Tween80 (1/4)), lh (20 mg/kg)
or 2k (15
mg/kg). Doses were selected based on acute toxicity studies of lh and 2k in
ICR mice (n =
2/group) showing that doses up to 30 mg/kg and 15 mg/kg, respectively, did not
cause greater
than 10% loss of body weight after 4 consecutive days of intraperitoneal
dosing.
[00502] In vivo antitumor activity [tumor growth inhibition (% TIC),
tumor growth
delay (T-C value), and tumor cell kill (total log cell kill)]. Evidence of
drug effect is described
by the following parameters: % TIC = [A tumor volume of treated group] / [A
tumor volume of
control group] x, 100%. The T-C values (tumor growth delay) were based on the
median time (in
days), required for the treatment (T) and the control group (C) tumors, to
reach a predetermined
size (600 mm3 in this study). These values were then used for the quantitation
of the tumor cell
kill following the equation: log cell kill = (T-C) / (3.32 ?ad). Td is the
tumor volume-doubling
time in days. In this study, we defined the doubling time required for the
tumor to increase from
300 to 600 mm3.
[00503] Rotarod test. ICR mice received training three times a day for
two days to enable
them to stay on the rotating rod for>120 seconds at 12 rpm. Mice were then
randomized by the
length of time that they could stay on the rotating rod and divided into 7-8
mice per group. lh at a
dose of 5 or 15 mg/kg in Captex200/Tween80 (1/4) was administered by
intraperitoneal injection.
Vinblastine at a dose of 0.5 mg/kg/day was used as a positive control under
the same conditions.
The rotarod test was performed twice a week. Treatment was stopped on day 31,
and post
observation was examined on weeks I, 2, and 4 after termination of the
treatment. The rod speed
was increased from 59 rpm to 40 rpm over a period of 5 min. Performance was
measured as the
length of time that a mouse could stay on the rotating rod.
[00504] In vivo drug resistance studies. At the end of the PC-3
xenograft studies, solid
tumors from control and lh treated (15 mg/kg) groups were removed and digested
with 0.1%
collagenase (Type I) and 50 mgJmL DNAse (Worthington Biochemical Corp.,
Freehold, N1).
Dispersed cells were plated in RPM! medium + 10% FBS and incubated at 37 C and
5% CO2 for
24 hr to allow attachment. The antiproliferative effects of lh were compared
to determine
whether tumor cells remaining in PC-3 xenografts retained sensitivity to drug.
The PC-3 cells
239
CA 3030689 2019-01-18
obtained from ATCC were used as in vitro control. Statistical analyses were
performed using
simple t-Test.
Results
[00505] Based on structure-activity relationship studies, three compounds
(Figure 28A)
were selected for biological characterization. While lh and 2k are highly
potent molecules with
low nanomolar cytotoxic properties, 21, which was rationally designed as a
potential metabolite
with improved solubility, had the least potent antiproliferative effects
(Table 21).
Table 21.1n vitro efficacy of compounds on prostate, melanoma and drug
resistant cell lines (n =
3, mean SE). Paciltaxel, vinblastine, and colchicine were used as positive
controls as previously
reported._
tcõõ *mu tutko
Cell bate Cell type SbikRT-H SbIART-F S5LART-011 Pa
rbtaxel ratblasritte Cob-Mane
LNCaP Prostate 25 4' 6 1' 10.3 9 1.7 * 0.2
1.1 0.1 16 4
PC-3 Pt ostate = 21 I .16 13 le VI 5 4.3 0.3 2.1
0.2 11 1
Du-145 Pi ostate 71 3' 12 116 14 5.1 *0.1 1St 1.1
10 I 2
FP('-1 Pi strife 43 5` 8 76 2 23 *03 1.1
0.4 20 1
B16-F1 Mamma 55 5' 43 21' 113 6 17* 2 4.7 0.7
29 5
A375 blebutoula 28 5' 33 14' 93 11 12 3 1.1
*0.2 20 3
0 VCAR-3 Ovation 35 ! 34 3 110 8 1.7 0.1 3_9 0.1. 17
1
411-VDR-RFS Ovati an 13 1 12. I 1 45 5 6263 634 532
t 57 1113 t 79
Resistance Factor 0.4 0.4 0.4 L333 149 65
SMART-H in Table 21 is lh ; SMART-F in Table 21 is 2k; and SMART-OH in Table
21 is 21.
SMARTs inhibit microtubule polymerization by binding to the colchicine
binding site on tubulin.
[00506] Bovine brain tubulin (>97% pure) was incubated with the
individual compounds
.20 (10 1.1M) to test their effect on tubulin polymerization (Figure 28B).
While lh and 2k inhibited
tubulin polymerization by 90%, 21 inhibited the polymerization by only 55%.
Previous studies
demonstrated a concentration-dependent inhibition of tubulin polymerization by
lh. In addition,
under the same experimental conditions, the 1050 for lb (4.23 itM) is similar
to that of colchicine
(4.91 RIV1). These data suggest that the compounds exhibit strong antitubulin
polymerization
240
CA 3030689 2019-01-18
activity that corresponds well with their cytotoxicity (Table 21). The ability
of the compounds to
compete for known binding sites on tubulin was determined using a novel MS
competitive
binding assay, which was developed in our laboratory. Three tubulin ligands,
corresponding to
the three binding sites on tubulin, colchicine, vinblastine, and paclitaxel
were used for these
competitive binding studies. It was found that, over a concentration range of
0.1-125 itM, lb
specifically competed with colchicine binding to tubulin, but it did not
compete with either
vinblastine or paclitaxel binding to tubulin (Figure 28C).
[00507] SMART compounds inhibit the growth of multidrug-resistant
cancer cell
lines.
[00508] The ability of the compounds to inhibit the growth of cancer
cell lines was
evaluated using the SRB assay. As shown in Table 21, the compounds inhibited
the growth of
several human cancer cell lines, including four prostate cancer cell lines,
and two melanoma cell
lines, with IC values in the low nanomolar range. Out of the three compounds,
21 was the least
potent (IC50 76-116 nM). 2k exhibited the best antiproliferative effect with
IC50 values between 6
and 43 nM in prostate cancer and melanoma cell lines. In addition, the effect
of the compounds in
the OVCAR-8 and NCl/ADR-RES cell lines was also evaluated (Table 21). The
compounds
were equally potent against MDR cell (NCI-ADR-RES) and the parent cell line
(OVCAR-8).
Paclitaxel, vinblastine, and colchicine exhibited relative resistance values
of 1333, 149, and 65
times, respectively (Table 21). These data indicate that the compounds
circumvent P-gp-
mediated drug resistance.
- [00509] SMART compounds arrest PC-3 (prostate) and A375 (melanoma)
cells in
G2/M phase of cell cycle and induce cell apoptosis. PC-3 and A375 cells were
exposed to 10,
50, 200, and 1000 nM of the compounds for 24 h. Treatment with the SMART
compounds
resulted in concentration-dependent accumulation of both PC-3 and A375 cells
in the G2/M
phase with concomitant decreases in the percentage of cells in GO/G1 phase
(Figures 29A and
29B). The proportion of cells in G2/M phase significantly increased when
treated with 50 to 200
nM of lb, 2k, 21. Apoptosis was then examined by measuring the level of
cytoplasmic DNA-
histone complexes in PC-3 and A375 cells after 24 h treatment. Increasing
concentration of the
SMART compounds increased the level of cytoplasmic DNA-histone complexes in PC-
3 and
A375 cells (Figure 29C). The effect was more pronounced in A375 cells than PC-
3 cells, but
241
CA 3030689 2019-01-18
apoptosis was evident in both cell types. lh and 2k induced moderate apoptosis
at a
concentration of 50 nM, while 21 induced apoptosis only at concentrations
greater than or equal
to 200 nM,
[00510] In vivo PK profile of SMART compounds. A single dose bolus of
each
compound (15 mg/kg) was administered by tail vein injection to ICR mice to
characterize their
pharmacokinetics (Figure 30A). lh and 2k exhibited similar PK properties, but
21 exhibited
slightly greater AUC than lh and 2k indicative of a lower clearance for 21
(Table 22). 21 also had
2-3 times higher Vss than that of lh and 2k. The clearance values for all
three compounds were
equal to or higher than 90 mUmin/kg, the hepatic blood flow rate in mice,
suggesting that in
addition to hepatic removal, other degradation routes may be involved in the
elimination of the
compounds. The pharmacokinetic's of 111 and 2k (2.5 mg/kg) were also examined
in rats (Figure
30B). Interestingly, low clearance values and hepatic extraction rates were
obtained by both
compounds, suggesting that these compounds exhibit species differences in
clearance. In rats, lh
exhibited favorable pharrnacokinetic properties, which are low clearance (6
mllmin/kg),
moderate volume of distribution (7.6 Ukg), long half-life (24 hr), and high
exposure (AUC, 5.8
hr*m/mL) (Table 22) when administered iv.
25
35
242
CA 3030689 2019-01-18
Table 22. Pharmacolcinetic parameters of SMART compounds. SMARTs were
administrated 15 mg/kg and 2.5 mg/kg i.v. in mice and rats, respectively.
F
In vivo, pharinacoldnetic paraineters of SMART contpounds
Specks Parameter Unit SMART-H SMART-F SMART-OH
AUC larJtWtnL 1.9 2.2 7.6
Mice min 140 141 740
Likg 4.9 6.6 16.5
mLimiufkg 130 11.2 90
ATTC hr * 5.S 14 NA
Rats tin min 1431 2410 NA
Likg 7.6 34 NA
CL 6 11 NA
NA, not available
SMART-H in Table 22 is lh; SMART-F in Table 22 is 2k; and SMART-OH in Table 22
is 21.
[00511] SMART compounds inhibit prostate and melanoma xenografts growth
without neurotoxicity. Prostate cancer PC-3 and melanoma A375 tumors in mice
were allowed
to reach a volume of 150 mm3 and then tumor-bearing mice were treated with the
SMART
compounds. As shown in Figure 31A, tumor volumes in the control group
increased to 680 mm3
over the 21 day duration of the study. Tumor volumes in the lh treated group
increased to 370
mm3 (5 mg/kg treatment) and 176 mm3 (15 mg,/kg treatment) by day 21,
indicating strong anti-
tumor activity for this compound. Tumors in the 2k-treated animals increased
to 269 mm3 (5
mg/kg treatment) and 292 mm3 (15 mg/kg treatment), while animals in the 21(50
mg/kg) treated
group had tumors of 331 mm3 at day 21. This reduction in tumor volume reversed
upon
withdrawal of SMART compounds (data not shown). Table 23 summarized the in
vivo efficacy
(%T/C, T-C values, and log cell kill) of SMART compounds.
243
CA 3030689 2019-01-18
Table 23. /n vivo efficacy of SMART compounds (administered i.p.) on prostate
(PC-3),
melanoma (A375). %T/C, T-C value, and log cell kill are summarized. The
doubling time of
melanoma xenograft was 4.6 d. Vinblastine was used as the positive control. %
TIC 5_ 42% is
considered to be moderately active by National Cancer Institute criteria. NA,
not available.
Compound Dosage Xenop-aft % VC Median time to T-C Total lo
(inzikg) model reach 600 mm3. (days) cell kill
Vehicle NA Prostate 100 19 days NA NA
Vinblastiue 0.5 Prostate 29 NA NA NA
SMART-H 5 Prostate 29 NA NA NA
SMART-H 1.5 Prostate 4 NA NA NA
SMART-F 5 Prostate 21 NA NA NA
SMART-F 15 Prostate 24 NA NA NA
SMART-OH 50 Prostate 34 NA NA NA
Vehicle NA Melanoma 100 IS days NA NA
SMART-H 20 Melanoma 30 28 days 10 0.7
SMART-F 15 Melanoma 28 29 days 11 0.7
SMART-H in Table 23 is lh; SMART-F in Table 23 is 2k; and SMART-OH in Table 23
is 21.
[005121 lh tumor elicited %T/C = 29% and 4% at 5 and 15 mg/kg treatment
(all doses
were intraperitoneal (i.p.)), respectively, whereas, 2k elicited % T/C of 21%
and 24% at 5 and 15
mg/kg treatment, respectively. The high dose of 21(50 mg/kg) exhibited the
%T/C of 34%.
Vinblastine, the positive control, showed %T/C of 29% at day 22 in PC-3
xenografts (Figure
- 31B). Body weight measurements, to monitor toxicity, indicated that only
1 of 8 mice treated
with lh (15 mg/kg), and 2 out of 7 mice treated with 2k (15 mg/kg) lost more
than 15% body
weight. In addition to the antitumor effects of the compounds on PC-3 prostate
tumors, lh (20
mg/kg) and 2k (15 mg/kg) demonstrated a significant reduction of A375 tumors.
As shown in
Figure 31C, the tumor volumes of control group increased to 2183 mm3, whereas
the 14 volumes =
in lh and 2k treatment groups increased to 775 mm3 and 722 mm3, respectively.
lh and 2k
treatment evoked %T/C of 28% and 29%, respectively. Rotarod tests were
performed to examine
244
CA 3030689 2019-01-18
the in vivo neurotoxic effects of 1h. Based on the result of in vivo efficacy
experiments, 5 or 15
mg/kg [i.p. administration, Captex200/Tween80 (1/4)] of lh was chosen to study
the effect on
motor coordination. A 0.5 mg/kg treatment with vinblastine was used as the
positive control
under the same conditions. As shown in Figure 31D, vinblastine gradually
reduced the time (in
seconds) that the mice could stay on the rotating rod, and attained
significance by days 27 and 31
(p < 0.05) compared to the vehicle group. However, no significant difference
was observed in the
lh treatment groups, suggesting that lh did not cause neurotoxicity in ICR
mice at doses that are
associated with antitumor effects.
[00513] lh did not develop drug-resistance in PC-3 tumor bearing mice.
We excised
the PC-3 tumors from nude mice after 21 days of treatment with vehicle (n = 3)
or 15 mg/kg lh
(n = 3). Solid tumors were digested and dispersed into cells as described in
the methods section.
PC-3 cell line from ATCC (American Type Culture Collection, Manassas, VA, USA)
was used
as a control. IC50 values were 29.1 1.1, 29.1 0.8, and 30.4 0.5 nM in PC-
3 cells from ATCC,
and dissociated cells from vehicle and lh treated tumors, respectively. These
data demonstrate
that lh did not induce drug-resistance in PC-3 tumors after 21 days of
continuous 111 treatment.
EXAMPLE 23
MOLECULAR MODELING
Methods
[00514] All molecular modeling studies were performed with Schrodinger
Molecular Modeling
. Suite 2008 (Schrodinger LLC, New York, NY), running on a Dell Linux
Workstation. Because the
site of ABI compounds are much closer to that of ABT-751, rather than DAMA-
colchichine, we
selected tubulin complex with ABT-751 (PDB code: 3KHC) as our modeling system.
ABIs were
built and prepared using the Ligprep module, and they were docked into the ABT-
751 site using the
Glide module in Schrodinger Suite. The best docking complexes were subject to
restricted molecular
dynamics to release any strains using Macromodel module with OPLS-2005
forcefield. The ligand
and its surrounding residues within 15 A were allowed to move freely, while
residues outside the 15
A radius were kept rigid.
Results
[00515] Molecular modeling for binding ABI compounds in tubulin was
studied. Several
crystal structures of the ligand-tubulin complex are available in the PDB
databank, with the most
245
CA 3030689 2019-01-18
recent one from Dorleans et aL In general, the colchicine binding pocket
tolerates a variety of
molecular structures, which may indicate substantial conformation changes upon
ligand binding.
In fact, Dorleans et al. solved the crystal structures of both the empty
tubulin dimer and the
ligand-tubulin complex. They found that, without the presence of ligand, loop
7 (T7, residues
244-251, Figure 32) in the beta-monomer folds in to occupy the binding pocket,
while it flips out
upon ligand binding. The associated helix 7 (H7, residues 224-243) and helix 8
(H8, residues
252-260) were displaced upon ligand binding. It is conceivable that the extent
to which T7 is
displaced depends on the size of individual ligand. This flexibility presents
a significant
challenge to understand the precise binding modes for individual ligands
without solving actual
crystal structures. Nevertheless, careful analysis of the possible binding
modes could provide
some insights into the binding of different ligands.
[00516]
The binding modes of 12cb and llcb (stick model) are shown in Figure 32A and
32B. For comparison, the crystal structure complexes of ABT-751 and DAMA-
colchicine (wire
models) along with ABI-12eb/tubulin complex in Figure 32A is displayed. For
clarity, only the
related secondary structures forming the binding pocket in 13-tubulin are
shown in Figure 32A.
The overall structures of 12cb, ABT-751 and DAMA-colchicine overlapped very
well in the
. binding pocket. Several potential hydrogen bonding interactions
between compound 12cb and
tubulin were identified. The carbonyl group in 12cb was in sufficient
proximity to form two
hydrogen bond interactions with the backbone NH of Leu-252 in H8 and the
sidechain of Asp-
251 in T7 of the tubulin [3-monomer. The para-fluorine substituent in the C-
ring was close to the
sidechain of Cys241 in "17 and Tyr202 in S6, possibly forming one or two
hydrogen bonds. The
imidazole proton is very close and likely to form a hydrogen bond to Thr179 in
T5 loop (residues
173-182) of the tubulin a-monomer (Figure 32A). Together with the hydrophobic
interactions
provided by the aromatic rings, the likely formation of these hydrogen bonds
would contribute to
the high binding affinity to the tubulin dimer, resulting in high
antiproliferative potency.
[00517]
The binding mode of llcb will be conceivably less defined since two of the
three
aromatic rings may occupy the binding pocket in the 13-monomer while the third
ring may extend
toward the interface of the a/f3-monomers, similar to how the sidechain of
DAMA-colchicine binds.
Our modeling indicates that the protecting group likely extends to the tubulin
dimer interface, while
the A, C rings of llcb occupy similar binding pocket and orientation as 12cb
(Figure 32B). This
may explain the similar activity between the two compounds, even though llcb
has an extra ring
246
CA 3030689 2019-01-18
system. From the molecular modeling studies presented in Figures 32A and 32B,
the hydrogen
bond donor is likely to be the thiol group in Cys-241 in loop 7 of the(3-
subuint in a/(3-tubulin dimer.
[00518] The binding mode of ABI 12Th was modeled (not shown) and compared to
DAMA-
colchicine (see Figure 19 for structure of colchicine) in the a/13-tubulin
heterodimer. The overall
structure of 12fb and DAMA-cochicine overlapped very well. The p-fluoro phenyl
moiety overlaps
with the trimethoxylpheny moiety which is interacting with the T7 loop in the
(3-subunit. Similarly,
the p-chloro phenyl moiety occupies the other side of the pocket where the
seven-member ring of
the DAMA-cochicine is, with the chlorine atom occupying the pocket where the
methoxy moiety
interacts.
=
EXAMPLE 24
MI CR OTOBULE IMAGING
Materials and Methods
[00519] Cellomics Cytoskeleton rearrangement kit (Thermo Scientific,
Rockford, IL) was
used to get a visually appreciable proof of ABIs interacting with tubulin
inside the cells. WM-164
melanoma cells were treated with each compound for 18 h in duplicate using a
collagen-coated
96-well plate (Becton Dickinson Labware, Bedford, MA). Then cells were fixed
with 4%
paraformaldehyde (Thermo Scientific, Rockford, IL) and permeabilized using
permeabilization
buffer supply from the kit. Primary antibody for tubulin and fluorescence-
labeled secondary
antibody were subsequently added to the cells. Cell nuclei were stained by
DAN. Whole Cell
Stain Green was also applied to all cells. All images were acquired with an
Olympus IX71
inverted fluorescence microscope (Olympus Corp., Tokyo, Japan) with overlays
from separate
images of tubulin (red), nuclei (blue), and whole cells (green). For
comparison, paclitaxel,
colchicine and ABT-751, along with ABIs are included.
Results
[00520] Visual proof of ABIs interacting with tubulin inside the cells
was examined. The
mictotubule arrangement in human melanoma WM-164 cells upon treatment with
different
compounds is presented in Figure 33. The microtubule images clearly showed
that all five tested
compounds resulted in cytoskeleton rearrangement. There was a significant
difference between
paclitaxel and the other four compounds (colchicine, ABT-751, 12cb, and 12da).
Treatment with
247
CA 3030689 2019-01-18
paclitaxel resulted in a condensation of microtubules orderly lying around the
nuclei compared
with controls, consistent with its mechanisms of action for stabilizing
microtubules. On the
contrary, treatment with colchicine, ABT-75 I , 12cb, and 12da had similar
effects on
microtubules and resulted in some degree of microtubule fragmentation,
consistent with their
common mechanism of action for destabilizing microtubules. These results also
confirmed that
ABIs shared the same cellular target with colchicine and induced the same
cellular effect.
EXAMPLE 25
VASCULAR DISRUPTING ACTIVITY OF COMPOUNDS 17ya AND 55
Method
[00521] Cells. HUVECs (Human Umbilical Vein Endothelial Cells) were
cultured and
grown in EGM-2 BulletKit (Lonza, Cat No. CC-3162), which contains growth
supplements
including hydrocortisone, human fibroblast growth factor-basic with heparin
(hEGF-B), vascular
endothelial growth factor (VEGF), R3-insulin-like growth factor 1 (IGF-1),
ascorbic acid,
heparin, fetal bovine serum, human epidermal growth factor (hEGF), and GA-1000
(gentamicin
.. and amphotericin B) in Endothelial Cell Basal Medium-2. Cells between the
third and fifth
passages were used for experiments. PC-3 human prostate cancer cells and T47D
human breast
cancer cells were cultured in RPMI-1640 medium with 5% fetal bovine serum.
[00522] Cell growth inhibition studies. Cytotoxic or antiproliferative
activity of test
compounds was investigated in several cell lines using the sulforhodamine B
(SRB) assay.
Cultured cells were plated into 96-well plates and incubated in medium
containing different
concentrations of the test compounds for 24 h or 48 h. Cells were stained with
sulphorhodamine
B (SRB) solution. The optical density was determined at 540 nm on a microplate
reader (Dynex
Technologies, Chantilly, VA). Plots of percent inhibition of cell growth
versus drug
concentration were constructed, and the concentration that inhibited cell
growth by 50% relative
to the vehicle control (IC50) was determined by nonlinear least squares
regression using
VinNonlin software (Pharsight Corporation, Cary, NC).
[00523] Capillary formation and disruption assays. Capillary formation
assays were
performed in 96-well plates by plating 12,000 cells/well of HUVECs on a
Matrigel layer (BD
Biosciences). In order to evaluate the anti-capillary action, capillaries were
allowed to form over
a 16 h period before the addition of test compound or vehicle-control. In
addition, capillary
= 248
CA 3030689 2019-01-18
formation inhibitory effect of test compound was investigated by treating
HUVEC cells with test
compounds before capillary formation. Images were acquired immediately
following compound
addition, 5, 10, 15, and 25 h after exposure to test compound. Capillary
formation was quantified
by counting the number of tubes and nodes having at least three edges.
[00524] Endothelial monolayer permeability assay. The permeability of
an endothelial
cell monolayer was assessed in the transwell system. HUVECs were plated at 2 x
106 cells per
insert of 24 well plate in EGM-2 medium and incubated for 72 h to reach 100%
confluency. Test
compounds were diluted in EGM-2 medium and added to the upper chamber of the
apparatus.
Following 1, 2, and 4 h of incubation, the compounds were removed and 75
1.tg/mL FTTC-
conjugated dextran (MW 40,000) was added for 5 minutes. Fluorescent
measurements of the
lower chamber were taken after excitation at 485 nm and emission was measured
at 520 nm using
a BioTek Synergy 4 Microplate Reader.
Result
[00525] 17ya and 55 exhibited high antiproliferative activity against
endothelial cells.
17ya and 55 were evaluated for cytotoxic activity against growth
factor¨supplemented
endothelial cells and growth factor¨deprived endothelial cell cultures.
Combretastatin A-4 (CA4)
and doxorubicin were used as positive and negative control, respectively.
Compound 17ya
exhibited higher potency than compound 55 against actively proliferating
endothelial cells (Table
24 and Figure 35). Both 17ya and 55 exhibited selectivity for endothelial
cells showing lower
IC50 values compared to one of the prostate cancer cells. CA4, 17ya and 55
were 8, 5 and 3 times
more active against endothelial cells than against cancer cells, respectively,
while doxorubicin
was not specific to endothelial cells (Table 24 and Figure 35). However no
selectivity was
observed between quiescent and active endothelial cells with these compounds
(data not shown).
249
CA 3030689 2019-01-18
Table 24. Endothelial cell growth inhibition of 17ya and 55. N=3
CA4 Doxorubicin 17ya 55
Ho 0 OH 0
i * g
H300 "DR
NM
0
,0 0 OH 0.'4C:roil
F+3C0
,SCO H,C0
H,C0
NH, H3C0 OCH,
PC3 3.2 397.0 7.8 23.3
T47D 6.0 352.8 18.0 37.4
HUVEC 1.2 273.6 2.8 9.7
Selectivity
ratio*,
cancer 7.6 1.4 4.6 3.1
cells/
=
HUVEC
* To obtain the selectivity ratio between cancer cells and HUVEC cells, the
mean IC50 (nM)
values of test compounds in PC3 and T47D cells were used.
[00526] 17ya disrupts the formation of endothelial capillaries but does
not disrupt
preformed capillaries. The activity of 17ya was investigated on endothelial
cells engaged in
capillary tube formation in vitro. Endothelial cells were placed on a Matrigel
matrix and the
formation and construction of capillary tubes in the presence or absence of
compounds were
observed (CA4, doxorubicin, and 17ya).
[00527] To
avoid confusion between early stage of tube formation and disruption of tube
construction, HUVEC cells on matrix in the presence of drug treatment were
incubated for 15 It
Then disruption of capillary was determined by counting the number of tubes
and nodes in each
treatment group. On the other hand, to evaluate the effect of test compound in
preformed
capillaries, HUVEC cells on matrix were allowed to form capillary tube for 16
h and the
capillaries were treated with test compounds,
[00528] As a
result, the number of tubes and nodes was gradually decreased over time due
to deficiency or consumption of nutrient by HUVEC cells (Figure 36). This
trend was observed
in every drug treatment group (Figure 36). In order to examine the difference
between untreated
and pretreated capillaries 15 h incubation groups were compared (Figure 36).
250
CA 3030689 2019-01-18
[00529] Endothelial cells that were exposed to various concentrations
of 17ya (0 to 50 p.M)
plated on Matrigel matrix resulted in inhibition of tube formation in a dose
dependent manner.
17ya with approximate IC50 value of 5 nM in cell growth inhibition studies
inhibited more than
50% of tube formation compared to vehicle-control (Figure 37). 17ya at 10 nM
completely
inhibited the tube formation (Figure 37). However, in the preformed
capillaries, the 10 nM 17ya
treatment group did not disrupt the capillary structure by 15 h (Figure 36).
These results suggest
that 17ya inhibits the formation of endothelial capillaries significantly but
is less effective to
disrupt preformed capillaries. Similar result was observed in CA4 treatment
group (Figure 37).
However, doxorubicin did not affect the capillary construction at toxic
concentration.
[00530] 17ya and 55 increased the permeability of endothelial cell
monolayers.
Antitubulin agents could modify the integrity of endothelial cell layers
lining blood vessels by
targeting cytoskeleton of the endothelial cells. Thus, the vascular disruption
effect of antitubulin
agent is known to increase the permeability of blood vessel and thus could
lead to protein leakage
and high blood viscosity. This could result in reduction of blood flow,
causing subsequent tumor
death from hypoxia and nutrient deprivation.
[00531] The effect of 17ya and 55 was evaluated on vascular
permeability using in vitro
study using transwell system with confluent HUVEC monolayers. The change in
permeability by
test compound was measured by the leakage of dextran (MW 40,000) after 1, 2,
and 4 h of drug
treatment. CA4 was used as a positive control. CA4, 17ya, and 55 resulted in
increased
permeability and the effect was more pronounced at 1 h incubation (data was
not shown). 17ya
showed a potency similar to CA4 (Figure 38). Doxorubicin did not induce any
change in the
permeability of endothelial cell monolayer (Figure 38).
[00532] All of the features described herein (including any accompanying
claims, abstract
and drawings), and/or all of the steps of any method or process so disclosed,
may be combined
with any of the above aspects in any combination, except combinations where at
least some of
such features and/or steps are mutually exclusive. Although preferred
embodiments have been
depicted and described in detail herein, it will be apparent to those skilled
in the relevant art that
various modifications, additions, substitutions, and the like can be made
without departing from
the spirit of the invention and these are therefore considered to be within
the scope of the
invention as defined in the claims which follow.
251
CA 3030689 2019-01-18
